Phenolic Compound, Antioxidant, Antibacterial, and In Silico Studies of Extracts from the Aerial Parts of Lactuca saligna L.

Medicinal plants are considered a major source for discovering novel effective drugs. To our knowledge, no studies have reported the chemical composition and biological activities of Moroccan Lactuca saligna extracts. In this context, this study aims to characterize the polyphenolic compounds distributed in hydro-methanolic extracts of L. saligna and evaluate their antioxidant and antibacterial activities; in addition, in silico analysis based on molecular docking and ADMET was performed to predict the antibacterial activity of the identified phenolic compounds. Our results showed the identification of 29 among 30 detected phenolic compounds with an abundance of dicaffeoyltartaric acid, luteolin 7-glucoronide, 3,5-di-O-caffeoylquinic acid, and 5-caffeoylquinic acid with 472.77, 224.30, 196.79, and 171.74 mg/kg of dried extract, respectively. Additionally, antioxidant activity assessed by DPPH scavenging activity, ferric reducing antioxidant power (FRAP) assay, and ferrous ion-chelating (FIC) assay showed interesting antioxidant activity. Moreover, the results showed remarkable antibacterial activity against Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes with minimum inhibitory concentrations between 1.30 ± 0.31 and 10.41 ± 0.23 mg/mL. Furthermore, in silico analysis identified three compounds, including Apigenin 7-O-glucuronide, Quercetin-3-O-glucuronide, and 3-p-Coumaroylquinic acid as potent candidates for developing new antibacterial agents with acceptable pharmacokinetic properties. Hence, L. saligna can be considered a source of phytochemical compounds with remarkable activities, while further in vitro and in vivo studies are required to explore the main biological activities of this plant.

Chemical profile, antibacterial, antioxidant, and anisakicidal activities of Thymus zygis subsp. gracilis essential oil and its effect against Listeria monocytogenes.

In this work, the chemical composition of Thymus zygis subsp. gracilis collected from Ifrane, Morocco, along with the evaluation of the antibacterial, anti-biofilm of Listeria monocytogenes activities, larvicidal effect against L3 larvae of Anisakis, and antioxidant properties of its essential oil (TZG-EO), are reported. GC-MS and GC-FID analyses highlighted the presence of 84 volatile components and strong bactericidal and anti-biofilm activities against L. monocytogenes at a concentration of 0.02 % were demonstrated. Also, larvicidal effect against Anisakis larvae at concentrations of 0.01 and 0.005 % was attained leading to the death of all tested larvae within 4 h. The in situ antibacterial activity of TZG-EO (0.01 and 0.005 %) in smoked fish showed high efficiency against L. monocytogenes growth. TZG-EO could be used as potential antibacterial and larvicidal agents for fighting against foodborne pathogens and extending shelf life of food products.

Phenolic Compounds, Antioxidant and Antibacterial Activities of Extracts from Aerial Parts of Thymus zygis subsp. gracilis, Mentha suaveolens and Sideritis incana from Morocco.

Thymus zygis subsp. gracilis, Mentha suaveolens and Sideritis incana (Lamiaceae family) are well recognized for their medicinal, pharmaceutical and aromatic properties. The present study aimed to investigate for the first time the phenolic composition, the antioxidant and antibacterial properties of the extracts obtained from the aerial parts of these species collected from the Ifrane region of Morocco. The phenolic compounds were determined using high-performance liquid chromatography coupled to photodiode array and electrospray ionization mass spectrometry (HPLC-PDA-ESI/MS). The antioxidant activity was investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, reducing power and ferrous ion chelating assays. The antibacterial activity was evaluated against three Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium) and three Gram-positive bacteria (Staphylococcus aureus, Enterococcus faecalis and Listeria monocytogenes). A total of thirty-three, sixteen and thirteen phenolic compounds were positively identified and characterized in T. zygis subsp. gracilis, M. suaveolens and S. incana extracts, respectively. Among the extracts, M. suaveolens exhibited the highest antioxidant activity, followed by S. incana and T. zygis subsp. gracilis in both DPPH and reducing power assays; all extracts showed the lowest activity in the chelating assay. In addition, all extracts demonstrated a bactericidal effect against Gram-positive bacteria and bacteriostatic effect against Gram-negative bacteria. Therefore, the aerial parts of Moroccan T. zygis subsp. gracilis, M. suaveolens and S. incana might be considered as a valuable source of natural antioxidant and antibacterial agents with potential application in food and pharmaceutical industries.

Chemical composition, antioxidant capacity and antibacterial action of five Moroccan essential oils against Listeria monocytogenes and different serotypes of Salmonella enterica.

Essential oils (EOs) obtained from aromatic plants are rich in natural components with interesting antimicrobial effects. The aim of this study was to evaluate the chemical composition of EOs extracted from Origanum majorana (OM-EO), Mentha suaveolens (MS-EO), Rosmarinus officinalis (RO-EO), Salvia officinalis (SO-EO) and Mentha pulegium (MP-EO). Their antioxidant properties and antibacterial activity against Listeria monocytogenes and different serotypes of Salmonella enterica subsp. enterica were also studied. The EOs were extracted from plants by hydro-distillation and their chemical composition was determined by GC-MS. Terpinen-4-ol, 1,8-Cineole, Camphor, Limonene and Cinerone were the main chemical components found in OM-EO, RO-EO, SO-EO, MP-EO and MS-EO, respectively. To the best of our knowledge, Limonene and Cinerone were reported, for the first time, as the major components of MP-EO and MS-EO. Moreover, our results showed that MS-EO had the best antioxidant activity with an IC50 of 0.78 ± 0.05 mg/mL, EC50 of 1.53 ± 0.07 mg/mL, and RC50 of 0.98 ± 0.04 mg/mL, and the higher antibacterial activity using microdilution broth method with MIC of 0.5% for Salmonella and 0.25% for L. monocytogenes, while OM-EO had the best antibacterial activity using disc diffusion method (inhibition diameters ranged between 15.3 ± 0.3 mm and 18.5 ± 0.3 mm for Salmonella and between 20.1 ± 0.2 mm and 25.4 ± 0.4 mm for L. monocytogenes). However, OM-EO and MS-EO present the higher percentage of sub-lethally injured cells against S. enterica (5.50 ± 0.11%) and L. monocytogenes (5.23 ± 0.07%), respectively. From this study, we can conclude that the investigated EOs are rich in components with interesting antibacterial activity and they could be applied in food preparations as natural preservatives to extend the shelf life of food products and to inhibit the growth of food-borne pathogens.

Prevalence, characterization and antimicrobial resistance of Listeria monocytogenes isolated from beef meat in Meknes city, Morocco.

INTRODUCTION: Listeria monocytogenes is one of the most pathogenic bacteria related to the consumption of contaminated food. This study aims to determine the prevalence of L. monocytogenes in raw beef meat in Meknes city of Morocco, to evaluate its pathogenicity and resistance to antimicrobials. METHODS: During four seasons, a total of 140 samples were collected from supermarkets, butcheries and Souk (weekly traditional market). The PCR method was used to examine the presence of specific and virulence genes in the isolated strains, and also to identify their serotypes. The antimicrobial resistance was determined. RESULTS: The results show a prevalence of 7.14% which depends on retail sites and also on the season's variation. The majority of the strains were detected in butcheries (6 strains), and supermarkets (4 strains). Moreover, the majority of strains were detected during summer (50%). Concerning virulence genes, the seven researched genes were detected in 100% of isolated strains. The majority of strains were of the (1/2a, 1/2c, 3a and 3c) serogroup (70%), while two of them were of the (1/2b, 3b, 4b and 4d) serogroup (20%). All isolates were resistant to at least one antimicrobial, while three strains were resistant to nine tested antimicrobials. However, they were highly susceptible to amikacin, imipenem, gentamicin, sulfamethoxazole and chloramphenicol. CONCLUSIONS: According to results, isolated L. monocytogenes from analyzed beef meat shows a high level of pathogenicity and resistance to the most used antimicrobials in listeriosis therapy, which calls for the severe application of quality systems at the slaughterhouses and retail sites level.

Antimicrobial Resistance, Virulence Genes, and Genetic Diversity of Salmonella enterica Isolated from Sausages.

Salmonella is a major cause of morbidity and mortality in humans worldwide, and the infection with multidrug-resistant strains can cause severe diseases. This study was designed to evaluate the antimicrobial resistance, to detect the virulence genes, and to study the genetic diversity of isolated Salmonella strains using 16S rRNA sequences. For this, 34 Salmonella strains isolated from sausages were identified using biochemical and serological methods. Molecular tools were used to evaluate the presence of virulence genes (orgA, sitC, sipB, spiA, iroN, and sifA) using simplex and multiplex polymerase chain reaction (PCR) and to sequence 16S rRNA genes for phylogenetic analysis. The susceptibility to 24 selected antibiotics was also studied. The results of this study showed that all isolated Salmonella were positive for targeted virulence genes and were resistant to at least one antibiotic. However, the multidrug resistance was observed in 44% of isolated strains. The phylogenetic analysis of 16S rRNA sequences highlighted that Salmonella isolates were divided into 3 clusters and 3 sub-clusters, with a ≥98% similarity to Salmonella enterica species. From this study, we conclude that sausages are considered as a potential source of Salmonella, which could be a major risk to public health.

Antibiotic Susceptibility profile of Staphylococcus aureus isolated from sausages in Meknes, Morocco.

BACKGROUND AND AIMS: Staphylococcus aureus is one of the most common causes of foodborne disease worldwide, due to the consumption of food contaminated by their toxins. This study aimed to determine the prevalence and the antimicrobial resistance of S. aureus isolated from sausages in Meknes city of Morocco. MATERIALS AND METHODS: A total of 156 samples (Beef sausages, Turkey sausages, and Artisanal sausages "Merguez") were collected from different shopping sites (butchery, supermarket, street vendors, and weekly market "Souk") and used for the isolation of S. aureus. All the isolated strains were tested for their antimicrobials resistance to 16 antibiotics. RESULTS: Our results showed the presence of S. aureus in 63 samples (40.38%). Furthermore, the antimicrobial resistance study showed that 84.13% of isolated S. aureus were resistant to streptomycin, 76.20% to tetracycline, 42.86% to ampicillin, 41.27% to doxycycline, 38.1% to penicillin G, and 19.05% to chloramphenicol with the presence of 25 different phenotypic profiles. However, all isolated strains were sensitive to oxacillin, cefoxitin, gentamicin, and vancomycin. CONCLUSION: The findings of this study revealed consumption of sausages as a potential risk of foodborne poisonings because of its contamination with the multi-resistant strains of S. aureus. Moreover, this contamination is related to the season, sampling sites and the origin of the raw material.

Antimicrobial resistance and genetic diversity of Salmonella Infantis isolated from foods and human samples in Morocco.

OBJECTIVES: Genotyping of Salmonella strains is an important molecular tool to discriminate isolates and to improve epidemiological studies when an outbreak occurs. Among the DNA-based genotyping methods, pulsed-field gel electrophoresis (PFGE) is currently used to subtype Salmonella isolates. In this study, the feasibility of genotyping Salmonella enterica serotype Infantis strains using XbaI restriction enzyme was evaluated. Separation of restricted fragments was performed by PFGE. METHODS: To test the possibility of applying this methodology to epidemiological investigation, a collection of 26 Salmonella Infantis strains were tested for their susceptibility to 14 antimicrobial agents and were analysed by XbaI macrorestriction followed by PFGE. Detection of class 1 integrons as well as intI1 and blaTEM genes in resistant strains was also studied. RESULTS: Antimicrobial susceptibility testing showed that 84.6% (22/26) of Salmonella Infantis isolates were susceptible to all of the antimicrobials tested, whereas 7.7% (2/26) had low-level resistance to ß-lactams and harboured the blaTEM gene. A class 1 integron (0.8kb) and the intI1 gene (898bp) were detected in one Salmonella Infantis strain. However, five different PFGE profiles were defined by XbaI macrorestriction. CONCLUSIONS: The PFGE method demonstrated adequate typing ability and represents a powerful tool to discriminate the serotype Salmonella Infantis.

Occurrence of Clostridium perfringens in sausages sold in Meknes city, Morocco.

In Morocco, the consumption of meat products has experienced a sharp increase in recent years despite the presence of pathogenic bacteria due to hygiene failure. The present study was designed to determine the prevalence of Clostridium perfringens in sausages sold in Meknes city (Morocco) and to study the different factors affecting it contamination with this bacterium. To this end, 156 samples of sausages were taken in various shopping sites during one year from March 2014 to February 2015. The microbiological analysis was carried out using the specific medium for isolation and identification of C. perfringens. ANOVA test was used for Statistical analysis (p< 0.05). The results of this study showed the presence of C. perfringens in 77.56% (121 of 156) samples, with 88.88% (32 of 36) in street vendors, 79.16% (19 of 24) in a weekly market, 70.83% (51 of 72) in butchery and 62.5% (15 of 24) in a supermarket. The average rate was 2.42 Log CFU/g, with a minimum value of 0 CFU/g recorded in several outlets and a maximum value of 6.05 Log CFU/g recorded in butchery. This study reveals that the contamination rate of sausages with C. perfringens is related to the sausages origin, retail sites and seasonal variations related to temperature increase.

Prevalence, molecular and antimicrobial resistance of Salmonella isolated from sausages in Meknes, Morocco.

Salmonella is among the most important food borne pathogens worldwide contaminating a wide range of animal products including meat products. The aims of this study go through two steps: The first step is to estimate the proportion of sausages products contaminated with Salmonella in Meknes city (Morocco), which were collected from various shopping sites: butchery, street vendors, supermarket and souk (Weekly market combines the population of the small villages around Meknes city). The second one is to identify serovars, to determine the antimicrobials resistance patterns of isolates and to detect the invA and spvC genes. 34 (21.79%) Salmonella were isolated, recovered 4 serogroups and 12 serotypes. The most prevalent serotypes were Salmonella Corvallis (23.53%) and Salmonella Kentucky (17.65%). All Salmonella isolates were tested for their susceptibility to 18 selected antimicrobials agents, of which 100% were resistant to at least one antimicrobial, 85.30% (29/34) were resistant to two or more antimicrobials and 44.12% (15/34) were resistant to at least three antimicrobials. All Salmonella are resistant to ampicillin, 76.47% to streptomycin, 20.59% to sulfonamides, 17.65% to Tetracycline and 11.77% to Ofloxacin. The "ACSSuT" penta-resistance pattern was observed in tow of the Salmonella Typhimurium strains. In addition, this study showed that all Salmonella strains (34) were positive for invasion gene invA and negative for the virulence gene spvC.