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Objective: We aimed to estimate the prevalence of Staphylococcus aureus producing Panton–Valentine leucocidin (PVL) isolated from children diagnosed with osteoarticular infections (OAIs), and to examine risk factors and clinical features. Methods: This prospective study was conducted from January 2017 to December 2018. All hospitalised children diagnosed with S. aureus OAI are included. Blood cultures, articular fluids, synovial tissues and/or bone fragments were collected for bacteriological culture. Antimicrobial susceptibility tests were determined by disk diffusion method. Genes encoding methicillin resistance (mecA) and PVL virulence factors (luk-S-PV and luk-F-PV) were detected by multiplex polymerase chain reaction. The demographic, clinical, laboratory, radiographic and clinical features were reviewed prospectively from medical records. Results: A total of 37 children with S. aureus OAIs were included, 46% of them have PVL-positive infection and 70.6% were male. The mean age was 8.12 years (±4.57), and almost were from rural settings (76.5%). Children with Staphylococcus aureus producing Panton–Valentine leucocidin (SA-PVL) were significantly associated with type of infection (P = 0.005), location of infection (P = 0.037) and abnormal X-ray (P = 0.029). All strains SA-PVL+ are sensitive to methicillin, but one strain SA-PVL negative was methicillin-resistant S. aureus, confirmed by gene mecA positive. Conclusion: The prevalence of S. aureus infections producing PVL toxin was high in OAIs amongst Moroccan children, mainly due to methicillin-susceptible S. aureus. Type and location of infections and abnormal X-ray were significantly associated with SA-PVL. Routine diagnostic testing of PVL-SA, continuous epidemiological surveillance and multidisciplinary management of OAI is essential to prevent serious complications.

The aim of this research paper is to test the antistaphylococcal effect of 1,8-cineole, amoxicillin/clavulanic acid (AMC), and gentamicin, either separately or in combination against three Staphylococcus aureus strains isolated from patients suffering from osteomyelitis. This activity was tested in vitro by using the microdilution method and the checkerboard assay. The efficacy of these three antibacterial agents was then tested in vivo by using an experimental model of methicillin-resistant S. aureus osteomyelitis in rabbits. This efficacy was assessed after four days of treatment by counting the number of bacteria in the bone marrow. The obtained results in vitro showed that the combination of the AMC with gentamicin did not induce a synergistic effect, whereas the combination of the two antibiotics with 1,8-cineole did. This effect is stronger when AMC is combined with 1,8-cineole as a total synergistic effect was obtained on the three strains used (FIC ≤ 0.5). In vivo, a significant reduction was noted in the number of colonies in the bone marrow when rabbits were treated with AMC associated with either 1,8-cineole or gentamicin compared to rabbits treated with AMC, gentamicin, or 1,8-cineole alone. These results demonstrated that 1,8-cineole showed a synergistic effect in combination with both AMC and gentamicin, which offer possibilities for reducing antibiotic usage. Also, the AMC associated with 1,8-cineole could be used to treat MRSA osteomyelitis.

The aim of the present study was to determine the chemical composition and antioxidant activity of essential oils (EOs) from two Teucrium polium subspecies, to evaluate, also their antibacterial activities, against some nosocomial-bacteria. The phytochemical screening of essential oils was analyzed using gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry analysis (GC-MS). The antibacterial activities were assessed by disc diffusion method and minimal inhibitory concentration (MIC), against Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Citrobacter koseri and Acinetobacter baumannii) and Gram-positive bacteria Staphylococcus aureus. The antioxidant potential was evaluated in vitro by three assays, namely free radical scavenging activity against 1,1-diphenyl-2-picrylhydrzyl (DPPH), ferric reducing activity power (FRAP) and total antioxidant capacity. Twenty-six components were identified in the EO of Teucrium polium subsp. aurum representing. Its major component was Caryophyllene (19.13%) followed by γ-Muurolene (13.02%), τ-cadinol, (11.01%), α-Gurjunene (9.2%), Rosifoliol (8.79%), 3-Carene (7.04%). However, twenty two components were identified in the EO of T. polium subsp. polium. Its major components are 3-carene (16.49%), γ-Muurolene (14.03%), α-pinene (9.94%), α-phellandrene (6.93%) and Caryophyllene (7.51%). The antibacterial activity of both essential oils showed a higher activity against tested nosocomial bacteria especially against S. aureus and A. baumannii. The EO of T. polium subsp. aureum showed better antioxidant activity as measured by DPPH and FRAP assays with IC50 values of 3.7 ± 0.2 mg/ml and 2.31 ± 0.11 mg/ml, respectively. The total antioxidant capacity assay showed that T. polium subsp. aureum had a significant activity with value to 3308.27 mg equivalent to ascorbic acid/g of EO. The Moroccan T. polium essential oils could be exploited as an antimicrobial agent for the treatment of several infectious diseases caused by bacteria, especially, those who have developed resistance to conventional antibiotics. Keywords: Teucrium polium, GC-MS, Nosocomial infections, Antibacterial, Antioxidant

Background: Streptococcus pyogenes is responsible for a wide variety of diseases, including noninvasive and severe invasive infections. The emm gene encodes the M protein that is the virulence factor and immunological determinant of group A streptococci. Emm typing is the group A Streptococci (GAS) standard molecular typing method based on the amplification of the N terminal hypervariable region of the emm gene. Objectives: The aim of the present study was to determine the prevalence of GAS in children with pharyngitis and determine different types of emm gene in the GAS isolates using emm typing. Methods: The study was carried out over a period of 14 months (from February 2017 to March 2018). Throat samples were collected from cases aged ≤ 18 years with pharyngitis referring to a primary health care center in Fez, Morocco. GAS isolates were subjected to conventional tests to confirm species identification. Antimicrobial susceptibility testing was performed using the standard disk diffusion method. We researched emm gene by a polymerase chain reaction (PCR). Emm types were determined by a sequence-based protocol. Demographic and clinical data were recorded from each patient. Results: From a total of 177 throat samples, 11 isolates (6.2%) were identified as GAS in children with pharyngitis. Antibiotic sensitivity testing revealed that all the GAS isolates were sensitive to penicillin. The sequencing of the PCR products of the emm gene revealed that emm90 was the most obtained emm type (30,77%); while emm75 was the least type observed (7.7%). Conclusions: The emm90 is the most prevalent type detected from patients with tonsillitis. Penicillin and erythromycin are still the foremost effective antibiotics to treat GAS pharyngitis.

The presence and accumulation of antibiotics in the water environment has become emerging contaminants of concern causing disruption of ecosystems worldwide. We describehere the seasonal variation and the occurrence of antibiotic residues in Fez city surface water (Morocco). During one year between February 2014 and January 2015, 8 surface water samples were collected monthly. Quantification of the 7 antibiotics was performed byon-line Solid Phase Extraction (SPE) liquid chromatography–tandem mass spectrometry (LC–MS/MS). A total of 96 surface water samples were investigated and the results revealed that 100% of the sites were contaminated by at least one antibiotic. Amoxicillin had the highest concentration with maximum concentration (4107 ng L-1), followed by ciprofloxacin (1058 ng L-1) and sulfamethoxazole was the most widely detected (93%). Seasonal variation showed that the concentrationof antibiotics was higher in winter for trimethoprim (96 ng L-1), ciprofloxacin (438 ng L-1) and in summer for amoxicillin (1113 ng L-1), sulfamethoxazole (162 ng L-1) and erythromycin (47 ng L-1). The results from this research show that antibiotics are frequent contaminants in Fez city surface water. This is the first attempt to assess the occurrence of these 7 pharmaceutical residues in water samples in Fez Morocco.

This study was conducted in order to know the colonization rate of MDR enterobacteria in neonates during their hospitalization in neonatal intensive care unit (NICU). Furthermore, we investigated risk factors for potential colonization and molecular epidemiology of isolated resistant bacteria. This prospective study was carried out in the neonatology and intensive care unit department of the University Hospital of Fez (Morocco) from February 2013 to July 2015. All consecutive admitted newborns were screened for intestinal and nasal carriage of MDR enterobacteria at admission of the babies and during the hospitalization. During the study period, a total of 641 Enterobacteriaceae were isolated and Klebsiella pneumoniae was the predominated bacteria. Bacterial identification and antibiograms were performed according to the international standards. On admission, 455 newborns were screened. A median age of these newborns was 1 day with an extended 147 days and their average weight was 2612 ± 1023 grams. 22.4% of neonates were found colonized by an ESBL producing Enterobacteriaceae (ESBL-E), 8.7% by a carbapenemases producing Enterobacteriaceae (CPE). During hospitalization, 207 of newborns were included in the acquisition study. 59.4% of newborns acquired an ESBL-E during their stay, 12.5% has acquired CPE. The blaCTXM-15 gene was the most frequently detected (81.2%) among ESBL-E. While, all CPE has expressed the blaOXA-48 gene exclusively. Two risk factors have been significantly associated with MDR enterobacteria colonization at admission which are newborns admission from maternity of the university hospital (95% CI, 1.859–5.129, P = 0.000) and neurological distress (95% CI, 1.038 to 4.694, P = 0.040). During hospitalization, the none risk factor was significantly associated with the carriage of MDR-E. The high rate of colonization, the MDR enterobacteria and the resistance genes found represent good indicator of cross-transmission in the NICU. An active strategy to control the spread of MDR enterobacteria should be applied.

During the Covid-19 pandemic, healthcare workers were extremely vulnerable to infection with the virus and needed continuous protection. One of the most effective and widely used means of protection was the FFP2 respirator. Unfortunately, this crisis created a shortage of these masks, prompting hospitals to explore opportunities to reuse them after decontamination.An approach for assessing the filtration efficiency of decontaminated FFP2 masks has been proposed and applied to evaluate the possibilities of their safe reuse. The decontamination processes adopted are those based on moist heat or hydrogen peroxide. The approach introduces efficiency measures that define the filtration and protection capacity of the masks, which characterize both chemical and structural changes, and encompasses many techniques including scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA). The test protocol was applied to mask samples that had endured different decontamination cycles and the results of their efficiency measures were compared to brand-new masks’ performances.The main result was that chemical and structural characterization of the decontaminated masks have shown no substantial change or deformation of their filter media structures. Indeed, the respiratory resistance test has shown that the results of both the FFP2 masks that have undergone a hydrogen peroxide disinfection cycle or a steam autoclave cycle remained constant with a small variation of 10 Pa from the EN149 standard. The chemical characterization, on the other hand, has shown that the filter media of the decontaminated masks remains unchanged, with no detectable chemical derivatives in its constituents.

The present work was designed to search the possible antibacterial effect of the ethanolic extract of pollens from six botanical origins, Punica granatum, Quercus ilex, Centaurium erythraea, Coriandrum sativum, Ruta graveolens, and Citrus aurantium, against multidrug pathogenic bacteria, Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus. The content of phenolic compounds, flavones, and flavonols was measured. The antioxidant activities were evaluated using four assays: total antioxidant capacity, DPPH, ABTS, and reducing power. Antibacterial activity was studied using the agar disk diffusion method, and the MIC and MBC were determined. Results obtained showed a positive correlation between the antioxidant content of pollen extracts and the antibacterial capacity, Punica granatum and Quercus ilex pollen extracts were the most efficient against Gram-positive and Gram-negative bacteria, and Centaurium erythraea, Coriandrum sativum, and Ruta graveolens had a moderate effect, while Citrus aurantium had no antibacterial effect. It is concluded that pollens can be a good source of bioactive molecules that exhibit potent antioxidant effects and strong antibacterial activities.

The aim of this research paper is to test the antistaphylococcal effect of 1,8-cineole, amoxicillin/clavulanic acid (AMC), and gentamicin, either separately or in combination against three Staphylococcus aureus strains isolated from patients suffering from osteomyelitis. This activity was tested in vitro by using the microdilution method and the checkerboard assay. The efficacy of these three antibacterial agents was then tested in vivo by using an experimental model of methicillin-resistant S. aureus osteomyelitis in rabbits. This efficacy was assessed after four days of treatment by counting the number of bacteria in the bone marrow. The obtained results in vitro showed that the combination of the AMC with gentamicin did not induce a synergistic effect, whereas the combination of the two antibiotics with 1,8-cineole did. This effect is stronger when AMC is combined with 1,8-cineole as a total synergistic effect was obtained on the three strains used (FIC ≤ 0.5). In vivo, a significant reduction was noted in the number of colonies in the bone marrow when rabbits were treated with AMC associated with either 1,8-cineole or gentamicin compared to rabbits treated with AMC, gentamicin, or 1,8-cineole alone. These results demonstrated that 1,8-cineole showed a synergistic effect in combination with both AMC and gentamicin, which offer possibilities for reducing antibiotic usage. Also, the AMC associated with 1,8-cineole could be used to treat MRSA osteomyelitis.

Background. The microbiological risk of the hospital environment, including inert surfaces, medical devices, and equipment, represents a real problem. Objective. This study is aimed at demonstrating and assessing the antibacterial activity of three synthetic disinfectants classified as quaternary ammoniums on different bacterial strains (Gram-negative and Gram-positive like Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, Acinetobacter baumannii, and Staphylococcus aureus) isolated from the hospital environment. The reference strains included Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, and Pseudomonas aeruginosa ATCC 27853 used as negative control strains. Method. Three quaternary ammonium disinfectants were tested: DDN9® (0.5%) which contains didecylmethylpolyoxyethylammonium propionate as an active substance, spray (0.4%) containing quaternary ammonium compounds, and Phagosurf ND® (0.4%) with didecyldimethylammonium chloride. Their effect was evaluated using the disk diffusion technique and the broth dilution methods, allowing the Minimum Inhibitory Concentration (MIC) and then the Minimum Bactericidal Concentration (MBC). Result. Only the growth of Gram-positive bacteria and some strains of Gram-negative bacteria were inhibited by the three synthetic disinfectants. NDD9® demonstrated an antibacterial effect only against the Gram-positive strains (S. aureus and S. aureus ATCC 29213) with a MIC of 0.25 mg/ml. The disinfectant spray showed effect against all four strains including E. coli (9), S. aureus, E. coli ATCC 25922, and P. aeruginosa ATCC 27853 with an inhibitory concentration of 4 mg/ml, while the growth of S. aureus ATCC 29213 was inhibited at 2 mg/ml. The third disinfectant, Phagosurf ND®, inhibited only the growth of S. aureus ATCC 29213 at a MIC of 4 mg/ml. Conclusion. This study is the first here in Morocco to evaluate the bacterial activity of products intended for the control of the healthcare environment. The results obtained on the three disinfectants tested reveal an ineffectiveness against some isolated strains from the hospital environment.

Objectives The aim of this study was to investigate the faecal carriage and molecular epidemiology of extended-spectrum β-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. bla CTX-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 bla OXA-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 bla CTX-M-15 and bla TEM , but not bla SHV . These genes were carried by a high-molecular-weight conjugative plasmid (ca. 125 kb). 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.

This study was conducted in order to assess the acquisition rate of Acinetobacter baumannii by newborn screening, on admission and during the discharge process of neonatal intensive care unit. (NICU). Furthermore, we investigated risk factors for potential colonization and molecular epidemiology of isolated resistant bacteria. This prospective study was conducted in the neonatal unit of Hassan II University Hospital of Fez from February 2013 to July 2015. During this period, all consecutive admitted neonates were screened for A. baumannii intestinal carriage, on admission and during the discharge process. Bacteriological and molecular tests were evaluated according to the international standards. This study examines the screening on admission of 455 newborns, 59% of whom were male. The average gestational age and birth weight were 35.2 weeks and 2612.1 g respectively. In total, 277 patients were included in the acquisition study on admission. The prevalence of multi-drug resistant (MDR) A. baumannii strain carriage was 6.5%, while the acquisition rate during the hospital recovery was 13.7%. In this study, 68 MDR A. baumannii isolates were collected. The resistance rates to different antibiotic classes including, Ceftazidime, Gentamycin and Ciprofloxacin varied between 92 and 100%. Moreover, 13% of MDR A. baumannii isolates were carbapenemase producers and 88% harbored blaOXA-23 gene. On admission, three risk factors were significantly associated with A. baumannii colonization: age (OR, 2.803; IC95%, 1.191-6.596; P = 0.01), gender (OR, 0.382; IC95%, 0.158-0.921; P = 0.03) and the delivery birth at the Maternity of University Hospital (MUH), (OR, 0.196; IC95%, 0.071-0.540; P = 0.002). However during hospitalization, the only risk factor associated with acquisition of A. baumannii was the respiratory distress (OR, 2.270; IC95%, 1.055-4.881; P = 0.03). A high intestinal carriage rate of A. baumannii and multiple antibiotic resistance were found in our NICU. Thus, the spread of MDR A. baumannii should be monitored by an active surveillance strategy.

Resume Le nombre d’infections nosocomiales et leur resistance croissante aux antibiotiques sont une preoccupation mondiale et peuvent toutefois etre reduits par le respect des bonnes pratiques. Les objectifs principaux de ce travail ont consiste a evaluer le respect des precautions standard ainsi que la conception du centre d’hemodialyse et a apprecier leur evolution suite aux actions mises en place. Deux enquetes transversales ont ete realisees au centre d’hemodialyse de l’hopital Al Ghassani de la ville de Fes (Maroc), la premiere avant amenagement du centre, la seconde apres sensibilisation du personnel soignant et renovation du centre. Les donnees ont ete recueillies par observation et suivi de la population cible, sur des grilles comportant 58 questions. Une amelioration generale dans l’infrastructure du centre renove et une defaillance de la pratique de certaines precautions standard aussi bien par le personnel que par les patients ont ete mises en avant. Des efforts restent a poursuivre en termes de formation et de diffusion des recommandations, d’elaboration et de mise en place de procedures, de protocoles manquants et de fiches d’enregistrements.

INTRODUCTION: Water treatment systems are a critical variable in dialysis therapy. Rigorous control of hemodialysis water quality is particularly important in order to guarantee a better quality of life of the hemodialysis patients. The objective of the study was to evaluate the chemical, microbiological quality and antimicrobial resistance of bacteria isolated from water and dialysate in a public HD center. METHODS: Fifty five samples of water and dialysate were collected weekly over a period of 4 months. The samples were collected from 4 points in the distribution loop. The microbiological and chemical analyses were performed according to our national standards. Antimicrobial susceptibilities patterns of isolated bacteria were determined by disk diffusion method. RESULTS: The chemical and microbiological parameters in all dialysis water and dialysate samples are in accordance with national standards. However, 70 Gram-negative bacteria were identified: Pseudomonas sp, Ochrobactrum antropi and Burkholderia cepacia, isolated at 52.8%, 12.8% and 17% simultaneously. Fourteen per cent of the isolates were resistant to three or more antibiotics. All resistant bacteria belong to the genus of Pseudomonas, 80% were resistant to tetracycline and to co-trimoxazole, 30% to ceftazidime. No colistin and imipenem resistance was observed. CONCLUSION: To avoid a health risk due to bacterial contamination, an adequate system for water treatment, disinfection of the hemodialysis system and microbiological monitoring of the water and dialysate are necessary.

Staphylococcus aureus, which has its ecological niche in the anterior nares, has been shown to cause a variety of infectious diseases mainly for patients in hemodialysis units. We performed this study to evaluate the prevalence of nasal S. aureus carriage among hemodialysis outpatients, to determine the antimicrobial susceptibility of isolates, to characterize the virulence genes, and to identify associated risk factors. Nares swab specimens were obtained from 70 outpatients on hemodialysis between March and June 2010. Samples were plated immediately onto S. aureus specific media and pattern of antibacterial sensitivity was determined using disk diffusion method. Polymerase chain reaction was used to detect nuc, mecA, and genes encoding staphylococcal toxins. Medical record of patients was explored to determine S.aureus carriage risk factors. Nasal screening identified 42.9% S. aureus carriers with only one (3.3%) methicillin-resistant S. aureus isolate. Among the methicillin-susceptible S. aureus isolates, high rate of penicillin resistance (81.8%) has been detected. The identified risk factors were male gender and age ≤ 30 years. Research of virulence factors showed a high genetic diversity among the 30 S. aureus isolates. Twenty-one (70%) of them had at least one virulence gene, of which 3.3% were Panton-Valentine leukocidin (lukS/F-PV) genes. S. aureus carriage must be screened for at regular intervals in hemodialysis patients. Setting up a bacterial surveillance system is one of the strategies to understand the epidemiology of methicillin-resistant S. aureus, to guide local antibiotic policy and prevent spread of antibiotic-resistant S. aureus.

Staphylococcus aureus, which has its ecological niche in the anterior nares, has been shown to cause a variety of infectious diseases mainly for patients in hemodialysis units. We performed this study to evaluate the prevalence of nasal S. aureus carriage among hemodialysis outpatients, to determine the antimicrobial susceptibility of isolates, to characterize the virulence genes, and to identify associated risk factors. Nares swab specimens were obtained from 70 outpatients on hemodialysis between March and June 2010. Samples were plated immediately onto S. aureus specific media and pattern of antibacterial sensitivity was determined using disk diffusion method. Polymerase chain reaction was used to detect nuc, mecA, and genes encoding staphylococcal toxins. Medical record of patients was explored to determine S.aureus carriage risk factors. Nasal screening identified 42.9% S. aureus carriers with only one (3.3%) methicillin-resistant S. aureus isolate. Among the methicillin-susceptible S. aureus isolates, high rate of penicillin resistance (81.8%) has been detected. The identified risk factors were male gender and age ≤ 30 years. Research of virulence factors showed a high genetic diversity among the 30 S. aureus isolates. Twenty-one (70%) of them had at least one virulence gene, of which 3.3% were Panton-Valentine leukocidin (lukS/F-PV) genes. S. aureus carriage must be screened for at regular intervals in hemodialysis patients. Setting up a bacterial surveillance system is one of the strategies to understand the epidemiology of methicillin-resistant S. aureus, to guide local antibiotic policy and prevent spread of antibiotic-resistant S. aureus.