Illicium verum L. (Star Anise) Essential Oil: GC/MS Profile, Molecular Docking Study, In Silico ADME Profiling, Quorum Sensing, and Biofilm-Inhibiting Effect on Foodborne Bacteria.

Illicium verum, or star anise, has many uses ranging from culinary to religious. It has been used in the food industry since ancient times. The main purpose of this study was to determine the chemical composition, antibacterial, antibiofilm, and anti-quorum sensing activities of the essential oil (EO) obtained via hydro-distillation of the aerial parts of Illicium verum. Twenty-four components were identified representing 92.55% of the analyzed essential oil. (E)-anethole (83.68%), limonene (3.19%), and α-pinene (0.71%) were the main constituents of I. verum EO. The results show that the obtained EO was effective against eight bacterial strains to different degrees. Concerning the antibiofilm activity, trans-anethole was more effective against biofilm formation than the essential oil when tested using sub-inhibitory concentrations. The results of anti-swarming activity tested against P. aeruginosa PAO1 revealed that I. verum EO possesses more potent inhibitory effects on the swarming behavior of PAO1 when compared to trans-anethole, with the percentage reaching 38% at a concentration of 100 µg/mL. The ADME profiling of the identified phytocompounds confirmed their important pharmacokinetic and drug-likeness properties. The in silico study using a molecular docking approach revealed a high binding score between the identified compounds with known target enzymes involved in antibacterial and anti-quorum sensing (QS) activities. Overall, the obtained results suggest I. verum EO to be a potentially good antimicrobial agent to prevent food contamination with foodborne pathogenic bacteria.

GC/MS Profiling, Antibacterial, Anti-Quorum Sensing, and Antibiofilm Properties of Anethum graveolens L. Essential Oil: Molecular Docking Study and In-Silico ADME Profiling.

Anethum graveolens L. has been known as an aromatic, medicinal, and culinary herb since ancient times. The main purpose of this study was to determine the chemical composition, antibacterial, antibiofilm, and anti-quorum sensing activities of the essential oil (EO) obtained by hydro-distillation of the aerial parts. Twelve components were identified, representing 92.55% of the analyzed essential oil. Limonene (48.05%), carvone (37.94%), cis-dihydrocarvone (3.5%), and trans-carvone (1.07%) were the main identified constituents. Results showed that the obtained EO was effective against eight bacterial strains at different degrees. Concerning the antibiofilm activity, limonene was more effective against biofilm formation than the essential oil when tested using sub-inhibitory concentrations. The results of anti-swarming activity tested against P. aeruginosa PAO1 revealed that A. graveolens induced more potent inhibitory effects in the swarming behavior of the PAO1 strain when compared to limonene, with a percentage reaching 33.33% at a concentration of 100 µg/mL. The ADME profiling of the identified phytocompounds confirms their important pharmacokinetic and drug-like properties. The in-silico study using molecular docking approaches reveals a high binding score between the identified compounds and known target enzymes involved in antibacterial and anti-quorum sensing (QS) activities. Overall, the obtained results highlight the possible use of A. graveolens EO to prevent food contamination with foodborne pathogenic bacteria.

Genome-Wide Identification and Expression Profiling of Pathogenesis-Related Protein 1 (PR-1) Genes in Durum Wheat (Triticum durum Desf.).

Pathogen-related proteins (PRs) are diversified proteins with a low molecular weight implicated in plant response to biotic and abiotic stress as well in regulating different functions in plant maturation. Interestingly, no systematical study has been conducted in durum wheat (Triticum turgidum subsp. durum). In the present study, 12 PR-1 genes encoding a CAP superfamily domain were identified in the genome of Triticum turgidum subsp. durum, which is an important cereal, using in silico approaches. Additionally, phylogenetic analysis showed that the PR-1 genes were classified into three groups based on their isoelectric point and the conserved motif domain. Moreover, our analysis showed that most of the TdPR-1 proteins presented an N-terminal signal peptide. Expression patterns analysis showed that the PR-1 gene family presented temporal and spatial specificity and was induced by different abiotic stresses. This is the first report describing the genome-scale analysis of the durum wheat PR-1 gene family, and these data will help further study the roles of PR-1 genes during stress responses, leading to crop improvement.

Methicillin-Resistant Staphylococcus aurous: Epidemiology, Transmission and New Alternative Therapies: A Narrative Review.

Over the last decade, we were facing medical struggle by the emergence of multi-resistant bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). MRSA infections are still causing a growing global concern due to the rapid adaptive multidrug resistance to conventional antibiotics in human, community and veterinary medicine. Here we provide an overview about MRSA epidemiology, transmission and alternative potential treatments particularly new discovered phytochemicals with biological activity. In this narrative review, bibliographic data was collected from literature search databases: Google Scholar, web of science and PubMed/MEDLINE during recent years (2016 to 2021). MRSA is responsible of wide spectrum life threatening infections such us septicemia, endocarditis, and wound infections. It has epidemic potential in hospitals, that is responsible of most nosocomial infections leading to mortality and constitute a real burden for the healthcare systems. Effective preventive strategies for management of MRSA are highly required moreover, the identification and development of novel drugs or active biomolecules through phytochemicals are time challenging to face new resistant strains.

Development of Nanotechnology-Based Drug Delivery Systems for Controlling Clinical Multidrug-Resistant Staphylococcus aureus and Escherichia coli Associated with Aerobic Vaginitis.

The growing prevalence of resistance to antibiotics potentially makes Escherichia coli and Staphylococcus aureus serious pathogens, necessitating the development of new antimicrobial agents. We extracted crude biosurfactants from a potential probiotic Bacillus spp. to control pathogenic bacteria associated with aerobic vaginal infection. Using nanotechnology formulations, we developed nanoemulsions based on biosurfactants at different concentrations (1% and 3.33%). The results showed that these nanoemulsions were stable, with a weighted index of 0.3, and demonstrated broad-spectrum antibacterial activity against Escherichia coli and Staphylococcus aureus, with MICs ranging between 1.25 and 4 mg/mL. Additionally, the nanoemulsions exhibited interesting antibiofilm effects. All strains became more sensitive to the antibiotics to which they were resistant because of various biosurfactant formulations combined with antibiotics. Lower concentrations of BNE1% and 3.33% were still more efficient than the crude biosurfactants. Our findings demonstrated that the biosurfactant had a strong antibiofilm effect against all tested pathogens. This antibacterial effect can be explained by their ability to alter cell physiology such as cell hydrophobicity and membrane disintegration. Thus, we can conclude that the use of nanotechnology formulations has improved this effect, and the nanoemulsions developed in this study can be used as a potential anti-infectious therapy against multidrug-resistant bacterial strains of clinical origin.

Identification of a putative kinase interacting domain in the durum wheat catalase 1 (TdCAT1) protein.

Catalases are crucial antioxidant enzymes that regulate plants responses to different biotic and abiotic stresses. It has been previously shown that the activities of durum wheat catalase proteins (TdCAT1) were stimulated in the presence of divalent cations Mn2+, Mg2+, Fe2+, Zn2+, and Ca2+. In addition, TdCAT1s can interact with calmodulins in calcium-independent manner, and this interaction stimulates its catalytic activity in a calcium-dependent manner. Moreover, this activity is further enhanced by Mn2+ cations. The current study showed that wheat catalase presents different phosphorylation targets. Besides, we demonstrated that catalase is able to interact with Mitogen Activated Proteins kinases via a conserved domain. This interaction activates wheat catalase independently of its phosphorylation status but is more promoted by Mn2+, Fe2+ and Ca2+ divalent cations. Interestingly, we have demonstrated that durum wheat catalase activity is differentially regulated by Mitogen Activated Proteins kinases and Calmodulins in the presence of calcium. Moreover, the V0 of the reaction increase gradually following the increasing quantities of Mn2+ divalent cations. Such results have never been described before and suggest i) complex regulatory mechanisms exerted on wheat catalase, ii) divalent cations (Mn2+; Mg2+; Ca2+ and Fe2+) act as key cofactors in these regulatory mechanisms.

Genome-Wide Identification and Expression Analysis of Catalase Gene Families in Triticeae.

Aerobic metabolism in plants results in the production of hydrogen peroxide (H2O2), a significant and comparatively stable non-radical reactive oxygen species (ROS). H2O2 is a signaling molecule that regulates particular physiological and biological processes (the cell cycle, photosynthesis, plant growth and development, and plant responses to environmental challenges) at low concentrations. Plants may experience oxidative stress and ultimately die from cell death if excess H2O2 builds up. Triticum dicoccoides, Triticum urartu, and Triticum spelta are different ancient wheat species that present different interesting characteristics, and their importance is becoming more and more clear. In fact, due to their interesting nutritive health, flavor, and nutritional values, as well as their resistance to different parasites, the cultivation of these species is increasingly important. Thus, it is important to understand the mechanisms of plant tolerance to different biotic and abiotic stresses by studying different stress-induced gene families such as catalases (CAT), which are important H2O2-metabolizing enzymes found in plants. Here, we identified seven CAT-encoding genes (TdCATs) in Triticum dicoccoides, four genes in Triticum urartu (TuCATs), and eight genes in Triticum spelta (TsCATs). The accuracy of the newly identified wheat CAT gene members in different wheat genomes is confirmed by the gene structures, phylogenetic relationships, protein domains, and subcellular location analyses discussed in this article. In fact, our analysis showed that the identified genes harbor the following two conserved domains: a catalase domain (pfam00199) and a catalase-related domain (pfam06628). Phylogenetic analyses showed that the identified wheat CAT proteins were present in an analogous form in durum wheat and bread wheat. Moreover, the identified CAT proteins were located essentially in the peroxisome, as revealed by in silico analyses. Interestingly, analyses of CAT promoters in those species revealed the presence of different cis elements related to plant development, maturation, and plant responses to different environmental stresses. According to RT-qPCR, Triticum CAT genes showed distinctive expression designs in the studied organs and in response to different treatments (salt, heat, cold, mannitol, and ABA). This study completed a thorough analysis of the CAT genes in Triticeae, which advances our knowledge of CAT genes and establishes a framework for further functional analyses of the wheat gene family.

Identification and Expression Profiling of Two Saudi Arabia Catalase Genes from Wheat and Barley in Response to Abiotic and Hormonal Stresses.

Catalase is a crucial enzyme in antioxidant defense systems protecting eukaryotes from oxidative stress. These proteins are present in almost all living organisms and play important roles in controlling plant responses to biotic and abiotic stresses by catalyzing the decomposition of H2O2. Despite their importance, little is known about their expression in the majority of monocotyledonous species. Here, we isolated and characterized two novel catalase genes from Triticum turgidum and Hordeum vulgare, designated as TtCAT1 and HvCAT1, respectively. Phylogenetic analysis revealed that TtCAT1 and HvCAT1 presented 492 aa and shared an important identity with other catalase proteins belonging to subfamily 1. Using bioinformatic analysis, we predicted the 3D structure models of TtCAT1 and HvCAT1. Interestingly, analysis showed that the novel catalases harbor a peroxisomal targeting signal (PTS1) located at their C-terminus portion, as shown for other catalase proteins. In addition, this motif is responsible for the in silico peroxisomal localization of both proteins. Finally, RT-qPCR analysis showed that TtCAT1 and HvCAT1 are highly expressed in leaves in normal conditions but faintly in roots. Moreover, both genes are upregulated after the application of different stresses such as salt, osmotic, cold, heavy metal, and hormonal stresses. The positive responses of TtCAT1 and HvCAT1 to the various stimuli suggested that these proteins can help to protect both species against environmental stresses.

Comparative Study of Antibacterial, Antibiofilm, Antiswarming and Antiquorum Sensing Activities of Origanum vulgare Essential Oil and Terpinene-4-ol against Pathogenic Bacteria.

Essential oils from aromatic and medicinal plants have many bioactive compounds known for their important biological activities mainly their antibacterial effects. Here we evaluated qualitatively and quantitatively the biofilm formation capability of pathogenic bacterial strains (n = 8). Then, we investigated the antibacterial, antibiofilm, antiquorum-sensing, and antiswarming efficacy of Origanum vulgare essential oil (EO) and terpinene-4-ol. Our results revealed that EO exhibited a more potent inhibitory effect against the tested strains. While the terpinene-4-ol was found to be more effective against developed Staphylococcus aureus biofilm. Regarding the anti quorum-sensing activity, we noticed that O. vulgare displayed better inhibition percentages in violacein production even at a low concentration (MIC/4). Additionally, this EO showed better inhibition of Pseudomonas aeruginosa PAO1 migration in comparison with the terpinene-4-ol. Our findings revealed that using pure O. vulgare EO demonstrated better competitive effects against pathogenic bacteria with a different mode of action when compared to the terpinene-4-ol. Hence, exploration and development of efficient anti-infection agents from natural resources such as full EOs represent promising tools in anti-infective therapy.

The Activity of the Durum Wheat (Triticum durum L.) Catalase 1 (TdCAT1) Is Modulated by Calmodulin.

Plant catalases (CAT) are involved in the cellular scavenging of the reactive oxygen species during developmental processes and in response to abiotic and biotic stresses. However, little is known about the regulation of the CAT activity to ensure efficient antioxidant function. Using bioinformatic analyses, we showed that durum wheat catalase 1 (TdCAT1) harbors highly conserved cation-binding and calmodulin binding (CaMBD) domains which are localized at different positions of the protein. As a result, the catalytic activity of TdCAT1 is enhanced in vitro by the divalent cations Mn2+ and Fe2+ and to a lesser extent by Cu2+, Zn2+, and Mg2+. Moreover, the GST-pull down assays performed here revealed that TdCAT1 bind to the wheat CaM (TdCaM1.3) in a Ca2+-independent manner. Furthermore, the TdCaM1.3/Ca2+ complex is stimulated in a CaM-dose-dependent manner by the catalytic activity of TdCAT1, which is further increased in the presence of Mn2+ cations. The catalase activity of TdCAT1 is enhanced by various divalent cations and TdCaM1.3 in a Ca-dependent manner. Such effects are not reported so far and raise a possible role of CaM and cations in the function of CATs during cellular response to oxidative stress.

Control of Staphylococcus aureus methicillin resistant isolated from auricular infections using aqueous and methanolic extracts of Ephedra alata.

In the current study the potential use of aqueous and methanolic extracts of Ephedra alata aerial parts as biological control agent against pathogenic bacteria and especially Staphylococcus aureus methicillin resistant isolated from auricular infections was evaluated. Chemical tests and spectrophotometric methods were used for screening and quantification of phytochemicals. The assessment of the antioxidant activity was accomplished by DPPH and ABTS radicals scavenging assays. Extracts were evaluated for their antibacterial efficacy by diffusion and microdilution methods. Biofilm inhibition was tested using XTT assay and the cytotoxicity of extracts was carried out on Vero cell line. The GC-FID analysis revealed that E. alata was rich in unsatured fatty acids. In addition, the aqueous extract had the highest flavonoid and protein contents (30.82 mg QE /g dry extract and 98.92 mg BSAE/g dry extract respectively). However, the methanolic extract had the highest phenolic, sugars and tannins. The antioxidant activity demonstrated that the aqueous extract exhibited the strong potency (IC50 ranged between 0.001 and 0.002 mg/mL). Both extracts displayed antimicrobial activity on Gram negative and positive strains. They were effective against S. aureus isolated from auricular infections. The tested extracts were able to inhibit biofilm formation with concentration-dependent manner. Moreover, no cytotoxic effect on Vero cells line was demonstrated for the extracts. Overall, our findings highlight the potential use of E. alata extract as a novel source of bioactive molecules with antioxidant, antibacterial and antiobiofilm effects for the control of infectious disease especially those associated to S. aureus methicillin resistant.

Pathogenic Impacts of Bacillus cereus Strains on Crassostrea gigas.

Regarding the economic importance of bivalve farming, a great deal of interest has recently been devoted to studying the pathogenesis of infectious diseases of these mollusks to prepare for public health emergencies. Bacillus cereus is one of these pathogens; it is a ubiquitous soil bacterium responsible for many types of gastrointestinal diseases associated with food. This study was conducted to determine the pathogenic effect of B. cereus on Crassostrea gigas. This effect was studied by assessing hemocytes death using flow cytometry analysis. The results showed that only ∼15% of C. gigas were able to survive after B. cereus artificial infection with 108 CFU (colony-forming unit)/oyster. Evenly, the percentage of nonviable hemocytes gradually increased with the concentration of B. cereus, with a peak value of ∼40% after infection. Indeed, findings showed that this strain is harmful to C. gigas.

Culture conditions improvement of Crassostrea gigas using a potential probiotic Bacillus sp strain.

It is well demonstrated that some probiotics improve rearing water quality and thereby have beneficial effects on reared organisms. We conducted this study to determine the effect of Bacillus consortium on Crassostrea gigas reared in contemned seawater with indigo dye priory treated with Bacillus or no treated. This effect was studied by assessing hemocytes death using flow cytometry analysis. We found that the percentage of decolorization of indigo dye in polluted seawater in presence of C. gigas increased from 41% to 90% when using Bacillus consortium. In these conditions, the hemocytes mortality of reared C. gigas decreased from 87% to 56%. We have demonstrated also that seawater contemned with priory treated indigo with Bacillus consortium is less toxic than seawater contemned with the no treated indigo. The percentage of hemocytes death is 81% for the contemned seawater with indigo and 56% for no contemned seawater. This consortium shows a protector effect of C. gigas against Vibrio harveyi contemning reared seawater.

Effect of environmental stress on cell surface and membrane fatty acids of Lactobacillus plantarum.

Adhesion has been regarded as one of the basic features of probiotics. We undertake this study in the aim to give new insight about the change in cellular physiological state under heat and acid treatments of Lactobacillus plantarum. Different cell properties have been investigated such as adhesive ability to abiotic surfaces, the cell surface hydrophobicity and the fatty acids profiles. The results of cell surface properties and Gas chromatography analysis demonstrated a modification in term adhesive ability and fatty acid (FA) composition of the tested strain under stressful conditions. In fact, after the exposure of the strain to heat and acid treatments, an increase in the hydrophobicity level and the adhesion capacity on HeLa cells was shown. Our findings revealed that high temperature and low pH change the fatty acids profiles of the treated cells, especially the proportions of unsaturated and saturated fatty acid. In this context, our data revealed that the unsaturated FA-to-saturated FA ratio was increased significantly (P < 0.05) for stressed strains compared with control cells. The results of the present finding suggest that the tested strain have suffered changes like the modifications on bacterial membrane as a cellular response to survive the hard environmental conditions, allowing them to withstand harsh conditions and sudden environmental changes to survive under.

Glucomannan's protective effect on the virulence of Vibrio splendidus in pacific oyster.

We examine the effect of Glucomannan, extracted from Candida utilis yeast, on immune parameters and resistance to Vibrio splendidus of Crassostreagigas. Our results showed that Glucomannan was a successful anti-adhesive molecule; it exhibited a stronger inhibitory effect on adhesion of Vibrio splendidus in infected Crassostreagigas. Vibrio splendidus viable cells number declined after incubation with Glucomannan. Furthermore, the Glucomannan diet showed higher activity to trigger the immune response against bacteria. Glucomannan applications, in biological control of seafood associated pathogens can be an alternative solution, providing consumer with a product of good quality owing to the use of 40 non-toxic compounds. Based on our results, Glucomannan could be used as a bio-protective culture in oyster's depuration to prevent Vibrio splendidus growth.

Change in cell surface properties of Lactobacillus casei under heat shock treatment.

We undertake this study in the aim to give new insight about the change in cellular physiological state under heat shock treatment and probiotic strain screening procedure. Different cell properties have been studied like adhesive ability to biotic and abiotic surfaces, the cell surface hydrophobicity and the fatty acids profiles. Compared to the normal cells, the heated cells increased their adhesive ability to biotic surface. However, the adhesion to abiotic surface was decreased. The cell surface hydrophobicity of the heated strains showed a significant decrease (P < 0.05). Our data revealed that high temperature change the fatty acids profiles of the treated cells, especially the proportions of unsaturated and saturated fatty acid. In fact, the ratio of saturated to unsaturated fatty acids of the heated Lactobacillus casei cells was significantly higher than that of the control cells (P < 0.05). The present finding could firstly add new insight about the response of probiotic to stressful conditions, such us the important role of cell membrane, considered as the first main structure to be damaged by physicochemical stress, in stress resistance because of their composition that can change in adaptation to harsh conditions. Secondly, there is no relationship between changes in membrane composition and fluidity induced by heat shock treatment and adhesion to biotic and abiotic surface.