Epidemiology and risk factors for staphylococcal urinary tract infections in the Moroccan Casablanca area.

PURPOSE: This study aimed to ascertain the prevalence and risk factors for developing staphylococcal urinary tract infections (UTIs) in the Casablanca area of Morocco. METHODS: In Casablanca, Morocco, a retrospective evaluation of 772 UTIs patients was conducted between January 2020 and December 2022. The research included two groups of patients: those with staphylococcal UTIs and those without. Sex, age, chronic illnesses, antibiotic exposure, urinary catheterization, urological surgery, and UTIs history were the risk variables assessed. We employed a logistic regression model to identify the characteristics that were predictive of staphylococcal UTIs. RESULTS: Eight staphylococcal species were responsible for 16.84% of UTIs in 772 non-repeating individuals. Patients infected with S. saprophyticus (35.38%) were the most common, followed by those infected with S. epidermidis (24.61%), S. aureus (13.85%), and S. hemolyticus (10.78%). Multivariate logistic regression analysis revealed that male sex (95% CI: 0.261-0.563), immunosuppression and immunosuppressive treatments (95% CI: 0.0068-0.64), chronic diseases (95% CI: 0.407-0.965), previous UTIs (95% CI: 0.031-0.228), frequency of urination more than 8 times a day (95% CI:1.04-3.29), frequency of urination once or twice a day (95% CI: 1.05-2.39), and urinary catheterization (95% CI: 0.02-0.22) were the most likely predictors of staphylococcal UTIs. In addition, a larger proportion of patients with staphylococcal UTIs were made aware of the risk factors associated with staphylococcal UTIs (52.31%, χ2 = 4.82, = 0.014). CONCLUSIONS: This is the first global study to evaluate the predictive factors for acquiring UTIs caused by staphylococci. Monitoring these factors will enable medical authorities to devise effective strategies for managing UTIs and combating antibiotic resistance.

Prevalence and antibacterial resistance patterns of uropathogenic staphylococci in Casablanca, Morocco.

INTRODUCTION: The purpose of this research is to evaluate the resistance profile of uropathogenic staphylococci bacteria in Casablanca, Morocco. METHODOLOGY: In this retrospective cross-sectional research carried out from January 2017 to December 2020, isolation and identification were carried out according to the usual techniques in medical microbiology. Staphylococcus aureus isolates were confirmed by polymerase chain reaction (PCR) amplification of the nuc gene, and the antibiogram was performed according to the guidelines of the Antibiogram Committee of the French Society of Microbiology (CA-SFM 2021). The susceptibility of uropathogenic staphylococci to vancomycin was determined with broth microdilution following the recommendations of the Clinical and Laboratory Standards Institute. The mecA gene was tested on phenotypically cefoxitin-resistant S. aureus isolates by PCR. RESULTS: The prevalence of urinary tract infections (UTIs) was 18% (772/4374). UTIs were more common in females (n = 483, 63%) than males (n = 289, 37%). Among the Gram-positive bacteria isolated (198, 25.65%), the prevalence of staphylococci was (130/198, 65.66%). Among staphylococcal species identified, coagulase-negative staphylococci (CoNS) were more prevalent (112/130, 86.15%), and Staphylococcus saprophyticus was the most frequently isolated CoNS (46/112, 41.07%). Additionally, there were several S. aureus strains (18/130, 13.85%). Forty-four percent of S. aureus isolates (n = 8) were resistant to cefoxitin and also harboured the mecA gene. All S. aureus isolates were susceptible to linezolid, cotrimoxazole and vancomycin. CONCLUSIONS: The prevalence and antibacterial resistance patterns of uropathogenic staphylococci in this study, with a high percentage of methicillin resistance, require careful consideration of antimicrobial therapy for staphylococcal UTIs.

Evaluation of Bioremediation Potentiality of Bacillus mojavensis Isolated from Wastewater for the Elimination of Reactive Yellow 145 and Methyl Orange.

Textile industry waste has become one of the largest polluters in the world. In recent years, there has been a growing awareness of the need for sustainable and eco-friendly practices for the treatment of dye-laden effluents. Overall, this study highlights the potential of bioremediation as a sustainable solution for wastewater treatment. The Bacillus mojavensis isolated from wastewater and identified using 16S rRNA degraded reactive yellow 145 and methyl orange in 36 h of incubation, this decolorization was affected by pH, temperature, dye concentration, glucose concentration, source of nitrogen, type of dye, and agitation. Our study found that the optimal conditions for total decolorization of dyes were achieved by incubating B. mojavensis at 46 °C, pH 9, with 1 g/L of glucose and 2 g/L of peptone. The azoreductase activity, FT-IR analysis, and UV-visible spectrum before and after total decolorization indicated that it was a dye degradation rather than biosorption in surface Celle. In addition, the study of phytotoxicity show the metabolites of degradation are not phytotoxic in Lens esculenta seeds. In conclusion, our results suggest the use of this bacterium as an environmentally friendly and also cost-effective method, making it an attractive option for industries looking to reduce their environmental impact.

Detection of Carbapenemase Encoding Gene and Resistance to Cefiderocol in Hospital and Community eXtensive Drug Resistance and Carbapenem-Resistant Pseudomonas aeruginosa Strains in Morocco.

Pseudomonas aeruginosa (Pa) remains among clinically-significant Gram-negative species. The carbapenems are often the last resort for treating infections due to multidrug resistant isolates such as Pa. The carbapenems' efficacy is increasingly compromised by the emergence and the rapid spread of Pa carrying carbapenemases which represent a serious threat to public health. This study aimed to establish the resistance profile and to identify carbapenemase genes in isolates with imipenem resistant phenotypes. Among 134 Pa isolates collected both in the community (46) and hospital (88) from January 2021 to December 2021 in Morocco, 18 (8 were from the community and 10 from the hospital settings) were carbapenem resistant. The identification of these strains has been confirmed using matrix assisted laser desorption ionization-time of flight (MALDI-TOF). The antibiotic susceptibility testing against 16 antibiotics was carried out and interpreted according to the recommendations of the European Committee on Antimicrobial Susceptibility Testing (2021). The worrying antibiotics resistance profiles, which spread to cefiderocol for two isolates, were obtained for all isolates, which were eXtensive Drug Resistance showing highly resistant to all antibiotic categories tested, even to ceftolozane-tazobactam. Colistin (100% susceptible) and cefiderocol (88.88%) were the most active agents against carbapenem-resistant Pa (CRPa). Phenotypic detection by NP-CARBA and NG-CARBA tests of metallo­ß­lactamase (MßL) production was confirmed by PCR amplification and sequencing. Three CRPa isolates coharboring blaVIM-2-blaNDM-1 (two isolates) and blaVIM-2-blaIMP-8 (one isolate) genes were detected. In this study, we describe the coexistence of these MßL genes and the cefiderocol resistance in CRPa strains in Morocco. The alarming antibiotic resistance patterns of all these CRPa isolates and their resistance genes emphasize the importance of antimicrobial susceptibility testing in the choice of antibiotics for treating Pa infections.

New alternative therapeutic strategies against Pseudomonas aeruginosa, an opportunistic multi-resistant pathogen with a myriad of virulence factors.

Pseudomonas aeruginosa (PA) has emerged as a significant cause of Gram-negative infections, particularly in patients with impaired host defenses. It is one of the six ESKAPE pathogens that majorly cause severe nosocomial infections. In addition to biofilm formation, PA possesses various virulence factors. It can be life-threatening due to his remarkable capacity to resist antibiotics, either intrinsically, developing adaptative resistance, or following the acquisition of resistance genes. The situation worsens when these mechanisms co-exist, conferring worrying multi-resistant phenotypes. Therapeutic options are becoming limited, which has led to the development of new antibiotics and novel alternative therapeutic strategies that require the exploration of other therapeutic avenues. Although mostly at the preclinical stages, many recent studies have reported several innovative therapeutic technologies that have demonstrated pronounced effectiveness in fighting against drug-resistant Pa strains. This literature review aims to discuss the mechanism of pyocyanic bacillus resistance to antibiotics, highlight the current state of some novel antibiotics and combination therapies, and the new alternative therapeutic approaches for treating PA infections.

Detection of ESBLs and carbapenemases among Enterobacteriaceae isolated from diabetic foot infections in Ouargla, Algeria.

INTRODUCTION: The emergence and rapid spread of Enterobacteriaceae carrying extended spectrum beta-lactamases (ESBLs) and carbapenemases represent a great threat to clinical treatment due to their multi-drug resistance. This study investigated ESBLs and carbapenemases encoding genes in Enterobacteriaceae collected from diabetic foot infections (DFIs) in Ouargla, southern Algeria. METHODOLOGY: A total of 70 Enterobacteriaceae strains were recovered from 76 patients with DFI between February 2017 and April 2018. Antimicrobial susceptibility testing was performed using the disc diffusion method, and the presence of bla genes was detected using polymerase chain reaction (PCR) and DNA sequencing. The genetic transfer of the plasmids was carried out by conjugation using the broth mating method. RESULTS: The most common isolate was Proteus mirabilis, followed by Escherichia coli, Morganella morganii and Klebsiella pneumoniae. The prevalence of ESBL and carbapenemase-producing Enterobacteriaceae was 11.42% and 2.85 % respectively. Plasmid-mediated AmpC was detected in 5.71% isolates. Conjugation experiments showed the transferability of blaCTX-M-2. CONCLUSIONS: Our findings support the view that various pathogens found in DFIs differ from one part of the country to another. This study reports the first description of metallo-ß-lactamase NDM-5 producing Klebsiella pneumoniae clinical isolate in Algeria.

Antimicrobial and antioxidant activities of Streptomyces species from soils of three different cold sites in the Fez-Meknes region Morocco.

The increasing demand for new bioactive compounds to combat the evolution of multi-drug resistance (MDR) requires research on microorganisms in different environments in order to identify new potent molecules. In this study, initial screening regarding the antimicrobial activity of 44 Actinomycetes isolates isolated from three soil samples from three different extremely cold sites in Morocco was carried out. Primary and secondary screening were performed against Candida albicans ATCC 60,193, Escherichia coli ATCC 25,922, Staphylococcus aureus ATCC 25,923, Bacillus cereus ATCC 14,579, other clinical MDR bacteria, and thirteen phytopathogenic fungi. Based on the results obtained, 11 active isolates were selected for further study. The 11microbial isolates were identified based on morphological and biochemical characters and their molecular identification was performed using 16S rRNA sequence homology. The UV-visible analysis of dichloromethane extracts of the five Streptomyces sp. Strains that showed high antimicrobial and antioxidant (ABTS 35.8% and DPPH 25.6%) activities revealed the absence of polyene molecules. GC-MS analysis of the dichloromethane extract of E23-4 as the most active strain revealed the presence of 21 volatile compounds including Pyrrolopyrazine (98%) and Benzeneacetic acid (90%). In conclusion, we studied the isolation of new Streptomyces strains to produce new compounds with antimicrobial and antioxidant activities in a cold and microbiologically unexplored region of Morocco. Furthermore, this study has demonstrated a significant (P < 0.0001) positive correlation between total phenolic and flavonoid contents and antioxidant capacity, paving the way for the further characterization of these Streptomyces sp. isolates for their optimal use for anticancer, antioxidant, and antimicrobial purposes.

Environmental surveillance of Legionella pneumophila in hot water systems of hotels in Morocco.

OBJECTIVE: Environmental monitoring of Legionella in hot water systems of hotels in Morocco was performed during the period from January 2016 to April 2018. A total of 149 water samples from 118 different hotels were analyzed. METHODS: A total of 149 water samples from 118 different hotels were analyzed. Possible risk factors were prospectively recorded, and data were analyzed in connection with building and plumbing systems characteristics. Data about building and risk factors were collected through a questionnaire survey. RESULTS: Out of the 149 samples, 77(51.7%) were positive for L. pneumophila. Serological typing of the isolates revealed that 54 (70.1%) are L. pneumophila serogroup 2-15 and 23 (29.9%) are L. pneumophila serogroup 1. 56.8% of all buildings were colonized by L. pneumophila. Counts were over 1,000 CFU/L in 44%. Contamination was strongly correlated with temperature in the circulation, the age of the premise plumbing and the size of the building. CONCLUSIONS: The results showed a relevant exposure to L. pneumophila in the community and the identified risk factors can serve as indicators for risk assessment and relevant actions.

Whole genomic comparative analysis of Streptococcus pneumoniae serotype 1 isolates causing invasive and non-invasive infections among children under 5 years in Casablanca, Morocco.

BACKGROUND: Streptococcus pneumoniae serotype 1 remains a leading cause of invasive pneumococcal diseases, even in countries with PCV-10/PCV-13 vaccine implementation. The main objective of this study, which is part of the Pneumococcal African Genome project (PAGe), was to determine the phylogenetic relationships of serotype 1 isolates recovered from children patients in Casablanca (Morocco), compared to these from other African countries; and to investigate the contribution of accessory genes and recombination events to the genetic diversity of this serotype. RESULTS: The genome average size of the six-pneumococcus serotype 1 from Casablanca was 2,227,119 bp, and the average content of coding sequences was 2113, ranging from 2041 to 2161. Pangenome analysis of the 80 genomes used in this study revealed 1685 core genes and 1805 accessory genes. The phylogenetic tree based on core genes and the hierarchical bayesian clustering analysis revealed five sublineages with a phylogeographic structure by country. The Moroccan strains cluster in two different lineages, the five invasive strains clusters altogether in a divergent clade distantly related to the non-invasive strain, that cluster with all the serotype 1 genomes from Africa. CONCLUSIONS: The whole genome sequencing provides increased resolution analysis of the highly virulent serotype 1 in Casablanca, Morocco. Our results are concordant with previous works, showing that the phylogeography of S. pneumoniae serotype 1 is structured by country, and despite the small size (six isolates) of the Moroccan sample, our analysis shows the genetic cohesion of the Moroccan invasive isolates.

Antimicrobial Susceptibility Patterns of Legionella spp. Strains Isolated from Water Systems in Morocco.

Objective: Legionella is a waterborne pathogen that causes a severe form of pneumonia called Legionnaires' diseases, which is normally acquired by inhalation of aerosols containing Legionella originating from natural and man-made water systems. The aim of this study was to describe the level of antimicrobial susceptibility of environmental Legionella spp. strains to preferred and recommended therapeutic agents to treat Legionella disease. Methods: The minimum inhibitory concentrations (MICs) of 60 environmental Legionella spp. strains were tested using the broth dilution method. Susceptibility testing was performed for 12 antimicrobial agents: macrolides (erythromycin, azithromycin [AZI], and clarithromycin [CLA]), fluoroquinolones (ciprofloxacin, levofloxacin, moxifloxacin, and gemifloxacin), a ketolide (telithromycin), cefotaxime (CEF), tigecycline (TIG), doxycycline (DOX), and rifampicin (RIF). Results: All tested strains of Legionella spp. were inhibited by low concentrations of fluoroquinolones and macrolides. Regarding the macrolides, CLA was the most active antibiotic, and AZI was the least active. RIF was the most effective antibiotic against the isolates in vitro. All isolates were inhibited by the following antibiotics (in decreasing order of their MICs): DOX>CEF>TIG. Conclusions: No resistance against these drugs was detected, and all isolates were inhibited by low concentrations of the tested antibiotics. Susceptibility testing of environmental Legionella spp. isolates must be monitored often to detect and evaluate the possible development of antibiotic resistance.

Dissemination of carbapenem-resistant Acinetobacter baumannii strains carrying the bla GES, bla NDM and bla OXA23 in Morocco.

BACKGROUND: Acinetobacter baumannii is a microorganism which has been classified by the World Health Organization in the list of the bacterial strains that pose the biggest danger to human health. This study was performed to determine the susceptibility profile to carbapenems and to detect carbapenemases production in 111 A. baumannii isolates. Among these 30 are environmental isolates and 81 are from the three major hospitals in Morocco. METHODS: All strains of A. baumannii were tested against diverse antimicrobial agents (13 antibiotic drugs) by the agar diffusion test. Minimum inhibitory concentration (MIC) of imipenem on carbapenem-resistant strains (CRAB) was determined by the E-test technique. Simple phenotypic tests were used to detect carbapenemases and metallo-ß-lactamases (MBLs) production including the modified Hodge test, EDTA test, and the cloxacillin test. The presence of carbapenemases-encoding resistance genes of CRAB strains was examined using polymerase chain reaction (PCR). RESULTS: Carbapenem resistance was observed in 23 clinical Acinetobacter isolates showing dissemination of the multiresistance profile. Molecular biology techniques indicated that all these strains encoded the naturally occurring bla OXA-51-like gene and were proved as A. baumannii. The bla OXA-23 gene was detected in 16 strains (69.6%). The metallo-ß-lactamase bla NDM gene was detected in five isolates (21.7%). GES-type carbapenemases were found in 15 strains, the existence of three classes of carbapenemases (bla GES, bla NDM, and bla OXA-23) was detected in three strains, while none of the CRAB isolates contained the bla OXA-58, bla OXA-24, bla VIM, bla OXA-48 or bla KPC encoding genes. CONCLUSIONS: This study established baseline proof of three classes of carbapenemases producing A. baumannii in Morocco, showing the important role of surveillance in controlling their spread.

Antibacterial activity of cinnamon essential oils and their synergistic potential with antibiotics.

The objective of this study was to evaluate the antibacterial activity of Cinnamomum cassia (cinnamon) essential oil (EO) alone and in combination with some classical antibiotics against three multidrug-resistant bacteria, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, to search a possible synergy. The antibacterial activity of all tested compounds was determined by agar disc diffusion and minimum inhibitory concentration assays. The checkerboard method was used to quantify the efficacy of cinnamon EO in combination with these antibiotics. Fractional inhibitory concentrations were calculated and interpreted as synergy, addition, indifferent, or antagonism. A synergistic interaction was shown against S. aureus with the combination cinnamon EO and ampicillin or chloramphenicol and against E. coli when cinnamon EO was combined with chloramphenicol. However, the combination of cinnamon oil and streptomycin displayed additive effects against all bacteria stains. The combinations of cinnamon EO and antibiotics can be used as an alternative therapeutic application, which can decrease the minimum effective dose of the drugs, thus reducing their possible adverse effects and the costs of treatment.

Virulence and Antibiotic Resistance Patterns in E. coli, Morocco.

Of 28 non-duplicate isolates of Escherichia coli recovered from yellow-legged Larus michahellis in Morocco, 92.86% were resistant to more than three antibiotics and 71.4% were multidrug resistant. Phylogenetic group A was most predominant (57.14%), followed by B1 (18%), B2 (14.28%) and F (10.71%). One isolate was resistant to ertapenem and contained the blaOXA-48 gene. The plasmid-mediated quinolone resistance determinants were detected in nine isolates (aac(6')-Ib-cr, qnrS1, qnrB1). Thirteen isolates carried one of the Shiga toxin E. coli-associated genes: stx1 (n = 6), stx2 (n = 5) and eae (n = 2) genes. Our data support the idea that gull feces may create potential public health risk.

Characterization of uropathogenic ESBL-producing Escherichia coli isolated from hospitalized patients in western Algeria.

INTRODUCTION: The aim of this study is to assess the prevalence and molecular characterization of uropathogenic Extended spectrum ß-lactamases (ESBLs) producing Escherichia coli. METHODOLOGY: During 3 years, all hospitalized patients at the University-affiliated hospital of Tlemcen and presenting urinary tract infections caused by E. coli were considered as potential study participants. These E. coli isolates were examined phenotypically for ESBL production. ESBL strains were subjected to antimicrobial susceptibility testing and were investigated for the presence of plasmid mediated quinolone resistance genes, 16SrRNA methylase genes and virulence genes by using conventional PCR and DNA sequencing. The molecular characterization of ESBL strains was established by phylogenetic grouping method and ERIC-PCR. RESULTS: The overall prevalence of ESBL was 32.5%. The blaCTX-M-15 was the most frequently detected in ESBL isolates, followed by blaCTX-M-14, blaCTX-M-28, blaCTX-M-1 and blaSHV-12 respectively. The plasmid-mediated quinolone resistance genes were detected in the 15 ESBL strains with the aac(6')-Ib-cr gene was the most detected followed by qnrB1 and qnrA1 gene respectively. Among the 22 ESBL isolates resistant to gentamicin and amikacin, the 16SrRNA methylase genes were detected in 4 isolates. The sfa and pap virulent genes were founds in 26% and 22% of isolates receptively. The genotyping analysis of all strains revealed that almost were belonged to phylogenetic groups A1 and A0 and fourteen distinct clones. CONCLUSION: The emergence of uropathogenic ESBL isolates and the high rate of blaCTX-M are alarming in Algeria. Strict measure must be required to control the further spread of these strains in Algerian hospitals.

Analysis of amino acid motif of penicillin-binding proteins 1a, 2b, and 2x in invasive Streptococcus pneumoniae nonsusceptible to penicillin isolated from pediatric patients in Casablanca, Morocco.

OBJECTIVES: This study aimed to investigate the nature of the amino acid motifs found in PBPs of Streptococcus pneumoniae isolates in invasive diseases from pediatric patients at Casablanca, Morocco. Five penicillin-susceptible (PSSP), ten penicillin-intermediate (PISP), and fifteen penicillin-resistant S. pneumoniae (PRSP) were studied by PCR-RFLP and DNA sequencing of the pbp1a, - 2b, and - 2x genes. RESULTS: There were no changes in the conserved motifs of PBP1a, PBP2b and PBP2x for PSSP strains. Substitution close to PBP1a conserved motifs were found in all PRSP isolates and six/five PISP. Analysis of PBP2b showed that all but one of the 10 PISP strains and all PRSP had substitutions. Substitution close to PBP2x motifs showed that all but three of the 10 PISP strains and all PRSP had substitutions in tow conserved motifs. A total of 6, 11 and 10 genotypes were found after analysis of pbp1a, pbp2b, and pbp2x, respectively. The penicillin-nonsusceptible S. pneumoniae isolated in Casablanca share most amino acid substitutions of those reported worldwide, but they occurred among pneumococci with low level resistance to b-lactams.

Adhesion of Legionella pneumophila on glass and plumbing materials commonly used in domestic water systems.

We aimed to investigate the adhesion of Legionella pneumophila serogroup1 and L. pneumophila serogroup2-15 on glass, galvanized steel, stainless steel, copper, Polyvinyl chloride(PVC), Cross-linked polyethylene(PEX-c) and Polypropylene Random Copolymer(PPR). The surface physicochemical properties of both bacterial cells and materials were estimated through contact angle measurements. The roughness and surface topography of the materials were evaluated by Atomic Force Microscopy. The two L. pneumophila serogroups and plumbing materials showed a hydrophobic character, while glass surface was hydrophilic. All strains were adhered to all materials with the exception of copper. The result showed that the adhesion of both L. pneumophila sg1 and sg2-15 was systematically expressed with high intensity on galvanized steel followed by PVC, PEX-c, PPR, stainless steel and the low intensity on glass. The extent of adhesion is in correlation with the surface roughness and acid-bases interactions, while hydrophobicity seems to have no effect in adhesion intensity.

Molecular characterization of penicillin non-susceptible Streptococcus pneumoniae isolated before and after pneumococcal conjugate vaccine implementation in Casablanca, Morocco.

BACKGROUND: Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide, especially among children and the elderly. The ability to effectively treat pneumococcal infection has been compromised due to the acquisition of antibiotic resistance, particularly to ß-lactam drugs. This study aimed to describe the prevalence and molecular evolution of penicillin non-susceptible S. pneumoniae (PNSP) isolated from invasive diseases before and after pneumococcal conjugate vaccine implementation in Casablanca, Morocco. METHODS: Isolates were obtained from the Microbiology Laboratory of Ibn Rochd University Hospital Centre of Casablanca. Serogrouping was done by Pneumotest Kit and serotyping by the Quellung capsular swelling. Antibiotic susceptibility pattern was determined by disk diffusion and E-test methods. The PNSP were analyzed by pulsed-field gel electrophoresis (PFGE) and by genotyping of pbp1a, pbp2b, and pbp2x genes. RESULTS: A total of 361 S. pneumoniae isolates were collected from 2007 to 2014. Of these isolates, 58.7% were obtained before vaccination (2007-2010) and 41.3% after vaccination (2011-2014). Of the 361 isolates, 80 were PNSP (22.2%). Generally, the proportion of PNSP between pre- and post-vaccination periods were 31 and 13% (p = 0.009), respectively. The proportion of PNSP isolated from pediatric and adult (age > 14 years) patients decreased from 34.5 to 22.9% (p = 0.1) and from 17.7 to 10.2% (p = 0.1) before and after vaccine implementation, respectively. The leading serotypes of PNSP were 14 (33 vs. 57%) and 19A (18 vs. 14%) before and after vaccination among children. For adults, serotypes 19A (53%) and 23F (24%) were the dominant serotypes in the pre-vaccination period, while serotype 14 (22%) was the most prevalent after vaccination. There were 21 pbp genotypes in the pre-vaccination period vs. 12 for post-vaccination period. PFGE clustering showed six clusters of PNSP grouped into three clusters specific to pre-vaccination period (clusters I, II and III), two clusters specific to post-period (clusters V and VI) and a cluster (IV) that contained clones belonging to the two periods of vaccination. CONCLUSION: Our observations demonstrate a high degree of genetic diversity among PNSP. Genetic clustering among PNSP strains showed that they spread mainly by a restricted number of PNSP clones with vaccine serotypes. PFGE clustering combined with pbp genotyping revealed that vaccination can change the population structure of PNSP.

Occurrence of plasmid-mediated quinolone resistance and virulence genes in avian Escherichia coli isolates from Algeria.

INTRODUCTION: The emergence and spread of quinolone-resistant Escherichia coli in poultry products puts consumers at risk of exposure to the strains of E. coli that resist antibiotic treatment. The objective of this study was to define the prevalence and virulence potential of poultry-associated nalidixic acid (NAL)-resistant E. coli in the Annaba city, Algeria. METHODOLOGY: In total, 33 samples of retail chicken meat were purchased from various butcher shops and examined for bacterial contamination with NAL-resistant E. coli. These isolates were subjected to antimicrobial susceptibility testing and were also investigated for the presence of plasmid-mediated quinolone resistance (PMQR) genes and virulence genes using conventional polymerase chain reaction (PCR) and DNA sequencing. Phylogenetic grouping of the NAL-resistant E. coli isolates was determined by the conventional multiplex PCR method. RESULTS: Twenty-nine (87.8%) products yielded NAL-resistant E. coli. Antibiograms revealed that 96.55% of NAL-resistant E. coli isolates were multidrug resistant (MDR). Resistance was most frequently observed against sulfamethoxazole-trimethoprim (96.6%), tetracycline (96.6%), ciprofloxacin (72%), and amoxicillin (65.5%). Group A was the most prevalent phylogenetic group, followed by groups D, B1, and B2. The PMQR determinants were detected in three isolates with qnrB72 and qnrS1 type identified. Four (13.8%) isolates carried one of the Shiga toxin E. coli-associated genes stx1, stx2, and ehxA alleles. CONCLUSIONS: The high prevalence of NAL-resistant E. coli isolated from retail chicken meat with detection of MDR E. coli harboring Shiga toxin genes in this study gives a warning signal for possible occurrence of foodborne infections with failure in antibiotic treatment.

Genotypic characterization of quinolone resistant-Escherichia coli isolates from retail food in Morocco.

This study was conducted to assess the retail food as a possible vehicle for antimicrobial resistant, particularly quinolones resistant and pathogenic Escherichia coli. We determined the prevalence and characteristics of nalidixic acid (Nal) resistant E. coli isolates from diverse retail food samples. In all, 70 (28%) of 250 E. coli isolates studied were Nal-resistant E. coli and 91% of these were multi-drug resistant. Plasmid mediated quinolone resistance genes were identified in 32 isolates, including aac(6')-Ib-cr (n = 16), qnrS1 (n = 11) and qnrB19 (n = 7). Mutations in gyr A and par C genes were detected among 80% of the isolates, and the isolates showed substitution Ser83-Leu and Asp87-Asn in gyrA and Ser80-Ile in parC. In addition, three different gene cassettes were identified (aadA1, aadA7, aac(3)-Id) in 18%. Virulence-associated genes stx1, eae, sfa, hlyA and stx2 were found in six (8%), three (4%), two (3%), three (4%) and three (4%) isolates, respectively. E. coli isolates of phylogenetic group A were dominant (64%, 45/70). Pulsed field gel electrophoresis revealed none epidemiological relationship between these isolates. The results of this work report the higher frequency of Nal-resistant E. coli isolates from Moroccan retail food samples including MDR and pathogenic isolates.

Rectal carriage of extended-spectrum ß-lactamase- and carbapenemase-producing Enterobacteriaceae among hospitalised neonates in a neonatal intensive care unit in Fez, Morocco.

OBJECTIVES: The aim of this study was to investigate the faecal carriage and molecular epidemiology of extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae (ESBLE) isolated from rectal samples of neonates hospitalised in a neonatal intensive care unit (NICU) of a university hospital in Fez, Morocco. METHODS: From February-July 2013, all neonates hospitalised in the NICU were screened for ESBLE carriage at discharge. ESBLs were identified by double-disk synergy test, PCR and DNA sequencing analysis. ESBLE were analysed by pulsed-field gel electrophoresis (PFGE), and conjugation was performed by the broth mating method. RESULTS: In this study, 169 Enterobacteriaceae were collected from 164 neonates. The prevalence of faecal carriage of ESBLE was 58.0% (98/169), predominantly Klebsiella pneumoniae (65/98; 66.3%). A high rate of multiresistance in ESBLE was noted. blaCTX-M-1 group (78.5%) was the most frequent ESBL gene detected, and all isolates harboured the CTX-M-15 variant. The prevalence of carbapenemase-producing Enterobacteriaceae was 1.8%, and blaOXA-48 was the only gene found in these isolates. Sequencing revealed subgroups corresponding to bla(CTX-M-15,TEM-1,TEM-104,SHV-1,SHV-44,SHV-49andSHV-133) genes. Conjugation experiments showed the transferability of blaCTX-M-15 and blaTEM, but not blaSHV. These genes were carried by a high-molecular-weight conjugative plasmid (ca. 125kb). PFGE profiles demonstrated high clonal dissemination of ESBL-positive strains in the NICU. CONCLUSIONS: These results demonstrate the existence of high clonal transmission of ESBLE in a Moroccan NICU. This finding provides useful information to implement a screening policy for resistant Enterobacteriaceae among neonates hospitalised in this ward.