Antibacterial activity and mechanism of cinnamon essential oil against Escherichia coli and Staphylococcus aureus

Cinnamon essential oil (EO) exhibited effective antibacterial activity against foodborne spoilage and pathogenic bacteria in model systems using Escherichia coli and Staphylococcus. The minimum inhibition concentration (MIC) of cinnamon EO was similar for both bacteria (1.0 mg/ml) while the minimum bactericide concentration (MBC) were 4.0 mg/ml and 2.0 mg/ml for E. coli and Staphylococcus aureus. GC–MS analysis confirmed that cinnamaldehyde was the major constituent in cinnamon EO (92.40%). Much effort was focused on elucidating the mechanism of antibacterial action of cinnamon EO against E. coli and S. aureus by observing the changes of cell microstructure using scanning electron microscope, determination of cell permeability, membrane integrity and membrane potential. After adding cinnamon EO at MIC level, there were obvious changes in the morphology of bacteria cells indicating cell damage. When cinnamon EO were added at MBC levels, the cells were destroyed. Cinnamon EO led to leakage of small electrolytes, causing rapid increase in the electric conductivity of samples at the first few hours. The values for E. coli and S. aureus reached 60% and 79.4% respectively at 7 h. Moreover, the concentration of proteins and nucleic acids in cell suspension also rose with increased cinnamon EO. Bacterial metabolic activity was decreased 3–5 folds as reflected by the results of membrane potential. Overall, S. aureus was more susceptible to cinnamon EO than E. coli.     

The specific antibacterial effect of the Salvia oil nanoliposomes against Staphylococcus aureus biofilms on milk container

The Staphylococcus aureus (S. aureus) biofilm on container is the main source of microbial contamination in milk. In this study, the nanoliposomes encapsulating Salvia oil (SO) has been prepared. Based on the damage effect of pore-forming toxin on cell membrane, α-toxin secreted from S. aureus has been used to trigger the release of SO from nanoliposomes to achieve antibacterial effect on S. aureus biofilm on milk container. Firstly, the minimum biofilm eradication concentration (MBEC) and biofilms time-dependent killing of SO were tested. The results showed that the MBEC of SO against S. aureus biofilms was 0.2% and SO effectively eradicated the biofilms after treating for 4 h. Subsequently, SO was encapsulated into nanoliposomes in order to increase its stability. The particle size, poly dispersity index (PDI), zeta potential, pH, turbidity, and entrapment efficiency of SO nanoliposomes were analyzed systematically. Gas chromatography–mass spectrometry (GC–MS) has been utilized to observe the controlled release of SO form nanoliposomes incubated with S. aureus. The scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) images have also visually showed that SO nanoliposomes have high anti-biofilm activity against S. aureus biofilms on milk container.     

Chemical composition,antibacterial activity and mechanism of action of essential oil from seeds of fennel (Foeniculum vulgare Mill.)

Fennel (Foeniculum vulgare Mill.) is widely cultivated and used as a culinary spice. In this work, the chemical composition of the essential oil obtained by hydrodistillation of fennel seeds was analyzed by gas chromatography-mass spectrometry (GC–MS), and 28 components were identified. Trans-anethole (68.53%) and estragole (10.42%) were found to be the major components. The antibacterial activity, minimum inhibitory concentration (MIC), and minimum bactericide concentration (MBC) of essential oil against several food-borne pathogens were evaluated. The results showed that the gram positive and gram negative strains of bacteria had different sensitivities to essential oil of fennel seeds, the essential oil exhibited antibacterial activity against Staphylococcus albus, Bacillus subtilis, Salmonella typhimurium, Shigella dysenteriae and Escherichia coli according to the results of MIC and MBC. Among these bacteria, S. dysenteriae was the most sensitive to essential oil, showing the lowest MIC and MBC values of 0.125 and 0.25 mg/mL respectively. In addition, kill-time assay also showed that the essential oil had a significant effect on the growth rate of surviving S. dysenteriae. We concluded that the mechanism of action of the essential oil against S. dysenteriae might be described as essential oil acting on membrane integrity according to the results of the leakage of electrolytes, the losses of contents (proteins, reducing sugars and 260 nm absorbing materials) assays and electron microscopy observation.     

Antifungal activity of citral,octanal and α-terpineol against Geotrichum citri-aurantii

This study investigated the antifungal activity and potential antifungal mechanisms of three volatile compounds (i.e., citral, octanal, and α-terpineol) against Geotrichum citri-aurantii, one of the main postharvest pathogens in citrus. Results showed that the volatile compounds exhibited strong antifungal activity against the targeted pathogens, with minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 0.50 μL/mL and 1.00 μL/mL for citral, 0.50 μL/mL and 2.00 μL/mL for octanal, and 2.00 μL/mL and 4.00 μL/mL for α-terpineol. The volatile compounds alter the morphology of G. citri-aurantii hyphae by causing loss of cytoplasm content and distortion of the mycelia. The membrane permeability of the G. citri-aurantii increased with increasing concentrations of the three volatile compounds, as evidenced by cell constituent release, extracellular conductivity, and pH. Moreover, the volatile compounds induced a decrease in the total lipid content of G. citri-aurantii cells, indicating the destruction of cell membrane structures. These results suggest that the antifungal activity of citral, octanal, and α-terpineol against G. citri-aurantii could be attributed to the disruption of cell membrane integrity and leakage of cell components.     

Bactericidal effects of Cinnamon cassia oil against bovine mastitis bacterial pathogens

Organic food production is expanding rapidly. However, this industry is hampered by the lack of effective antimicrobial agents which can be used in organic food production. This study examined the antimicrobial activity of Cinnamon cassia oil against major pathogens causing bacterial bovine mastitis, its miscibility in milk and possible antimicrobial mechanisms. C. cassia oil had inhibitory activity against all tested pathogen isolates from bovine mastitis. We conducted disk diffusion assay and found that discs with 20 μL of 2% (v/v) C. cassia oil solution resulted in inhibition zones of 29.6, 19.1, 27.0, 33.3 and 30.7 mm for Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hyicus, Staphylococcus xylosus and Escherichia coli 29, respectively. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of C. cassia oil was 0.00625% and 0.025% (v/v) for S. hyicus, 0.025% and 0.10% (v/v) for E. coli 29, and 0.0125% and 0.05% for S. aureus, S. epidermidis and S. xylosus, respectively. We selected two common mastitis pathogens, a representative S. aureus isolate and E. coli 29 for further analyses. Based on time-kill assay in LB broth with 0.15% agar, 2MBC of C. cassia oil generated bactericidal effects on S. aureus and E. coli 29 within 30 min, and 4MBC caused 6 log reduction of S. aureus and E. coli 29 within 30 min. In milk, C. cassia oil at 4MBC reduced ∼6.0 Log₁₀ CFU/ml of S. aureus and E. coli 29 to undetectable level within 8 h. Using propidium iodide staining, we observed membrane damage on both S. aureus and E. coli 29 cells during incubation with C. cassia oil. In addition, C. cassia oil treatment at MIC impaired membrane integrity of E. coli and S. aureus, which was followed by a decrease in ATP synthesis. Bacterial extracellular signaling quorum sensing orchestrates important events related to bacterial pathogeneses through excreting autoinducer (AI). Sub-inhibitory concentration of C. cassia oil repressed AI-2, a universal signal molecule mediating quorum sensing, production in S. aureus and E. coli 29 isolates. Collectively, our data show that C. cassia oil provides an exciting potential to be used as an alternative antimicrobial for bovine mastitis in organic dairy farms.     

Effects of cinnamaldehyde on Escherichia coli and Staphylococcus aureus membrane

Gram-negative Escherichia coli (ATCC 8735) and Gram-positive Staphylococcus aureus (ATCC 3101) were selected as model bacteria to determine the antimicrobial mechanism of cinnamaldehyde. Several techniques were utilized to investigate the effects of cinnamaldehyde on food-borne bacterial membranes. The ultraviolet (UV) absorption and electrical conductivity of the culture supernatant were used to determine membrane integrity. β-Galactosidase activity was determined to detect inner membrane permeability. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were performed to observe bacterial morphology. Samples from both strains exposed to cinnamaldehyde showed higher UV absorptions, conductivity values, and β-Galactosidase activities compared with the control group and displayed a rapid rise trend. Thereafter, the values stabilized at a relatively steady state. SEM shows that treated E. coli and S. aureus cell samples exhibited rough cell membranes with particulate matter, and some of the S. aureus cells split due to deep wrinkle formation and distortion, unlike the control group. TEM shows that the bacteria treated with cinnamaldehyde exhibited numerous abnormalities, including cytoplasmic membrane separation from the cell wall, cell wall and cell membrane lysis, cytoplasmic content leakage, cytoplasmic content polarization, cell distortion, and cytoplasmic content condensation. These results indicate that bacterial cell morphology, membrane integrity, and permeability are damaged when the E. coli and S. aureus cells are exposed to the minimum inhibitory concentrations of cinnamaldehyde (0.31 mg/mL). In addition, the higher the cinnamaldehyde concentration, the more serious the bacterial membrane damage is.     

The specific antibacterial activity of liposome-encapsulated Clove oil and its application in tofu

In this study, the antibacterial activities of Clove oil and liposome-encapsulated Clove oil were investigated. First, the antibacterial activity of Clove oil demonstrated that the essential oil exhibited favorable antimicrobial activity for both Escherichia coli and Staphylococcus aureus. However, a setback of using Clove oil as a disinfectant is its low chemical stability. Then Clove oil was incorporated into a liposome formulation to increase its stability. The optimal polydispersity index (PDI) (0.196), Zeta potential (−24.5 mV) and entrapment efficiency (20.41%) of liposome were obtained at the concentration of Clove oil to 5.0 mg/mL. In addition, selective antimicrobial activity for S. aureus by utilizing pore-forming toxins (PFTs) to activate Clove oil release from liposome was observed. By contrast, liposome-encapsulated Clove oil has no effect on E. coli that doesn't secrete PFTs because antimicrobial component can't reach bacteria. Gas chromatography (GC) assay found that when liposome met S. aureus that secrete PFTs, PFTs would insert into the liposome membranes and form pores, through which the encapsulated Clove oil was released. Besides, liposome-encapsulated Clove oil exhibited efficient antimicrobial activity for S. aureus in tofu.     

An insight into the inhibitory activity of dihydromyricetin against Vibrio parahaemolyticus

The objective of this study is to investigate the antibacterial activity of dihydromyricetin (DMY) against Vibrio parahaemolyticus. The dilution method indicated that the minimum inhibitory concentration (MIC) of DMY against V. parahaemolyticus was at 0.625 mg/mL. The inhibitory effects of DMY against V. parahaemolyticus was further studied by analyzing cell morphology, cell injury, cell permeability, cell surface hydrophobicity (CSH) and antibacterial rate. The results showed bacterium cells are completely inactivated in a higher concentration (10 MIC). DMY treatment also lead to an increase in cell membrane permeability, cell injury as well as CSH. A good correlation between antibacterial rate and CSH was also observed. These findings indicated DMY could be used as a new alternative natural antibacterial agent for control pathogen growth in aquatic food.     

Antibacterial activity of essential oils and their main components enhanced by atmospheric RF plasma

The antibacterial activity of low concentration clove oil, sweet basil oil, and lime oil (5–20 μl ml⁻¹) and their main components (eugenol, β-ocimene, and d-limonene) at the same concentration was enhanced by atmospheric RF plasma at 20W and 40W for 10 min to effectively control the growth of Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus on chicken egg. Number of colony germination after plasma treatment was evaluated. Results showed that growth of E. coli, S. thyphimurium, and S. aureus on eggshell mixed with clove oil (10 μl ml⁻¹) or its main component (eugenol at 5 μl ml⁻¹) was completely inhibited after using plasma treatment at 40W. Without the plasma treatment, all essential oils and all their main components at concentrations of 5–20 μl ml⁻¹ could not inhibit bacterial growth. Thus, this study has demonstrated the good potential of using atmospheric RF plasma treatment to enhance the antibacterial activity of essential oil at a relatively low concentration.     

Essential oil composition and antimicrobial activity of Sphallerocarpus gracilis seeds against selected food-related bacteria

Sphallerocarpus gracilis from China is a little-investigated edible and medicinal plant. In the present study, the essential oil composition from S. gracilis seeds was investigated by GC and GC–MS. A total of 34 compounds representing 94.69% of the essential oil were tentatively identified. The main constituents were p-cymene (17.42%), γ-terpinene (25.58%) and α-asarone (33.12%). The antimicrobial activity, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) of the essential oil from S. gracilis seeds were evaluated against eight Gram-positive bacteria (Listeria monocytogenes, Staphylococcus aureus, Staphylococcus epidermidis, Micrococcus luteus, Bacillus megaterium, Bacillus cereus, Bacillus coagulans and Bacillus subtilis), four Gram-negative bacteria (Salmonella enteritidis, Salmonella typhimurium, Klebsiella pneumoniae and Escherichia coli) and one fungus (Aspergillus niger). Results revealed that the essential oil from S. gracilis seeds exhibited significant in vitro antimicrobial property. Among all the tested microorganisms, the essential oil showed the strongest inhibitory effect against K. pneumoniae, whereas no inhibitory effect was found against L. monocytogenes and A. niger. Additionally, scanning electron microscopy (SEM) was used to observe morphological changes of bacteria treated with the essential oil from S. gracilis seeds. SEM observations confirmed the physical damage and considerable morphological alteration to the tested microorganisms treated with the essential oil. The data of this study suggests that the essential oil from S. gracilis seeds has great potential for application as a natural antimicrobial agent to preserve food.     

Citral inhibits mycelial growth of Penicillium italicum by a membrane damage mechanism

This study investigated the anti-fungal activity and potential anti-fungal mechanisms of citral against Penicillium italicum, one of the main citrus postharvest pathogens. Results showed that citral exhibited strong anti-fungal activity against the test pathogen, with minimum inhibitory concentration and minimum fungicidal concentration of 0.50 and 1.00 μL/mL, respectively. Citral could evidently alter the morphology of P. italicum hyphae by causing loss of cytoplasm and distortion of mycelia. The membrane permeability of the P. italicum increased with increasing concentrations of citral, as evidenced by the release of cell constituents, extracellular pH, and leakage of potassium ions. Moreover, citral could induce a decrease in total lipid and ergosterol contents of P. italicum cells, indicating the disruption of membrane integrity. These results suggest that the anti-fungal activity of citral against P. italicum can be attributed to the disruption of the cell membrane integrity and membrane permeability.     

Further analysis of the antimicrobial activity of α-phenylseleno citronellal and α-phenylseleno citronellol

In this work we performed studies on the antimicrobial activities of α-phenylseleno citronellal and α-phenylseleno citronellol, two new selenium-containing citronella oil derivatives. The presence of phenylselenium group in the citronellal molecule increased antimicrobial activity, as compared to a natural, unsubstituted terpene. We assessed the antimicrobial activity of each compound against three important species of foodborne pathogenic bacteria: Listeria monocytogenes, Staphylococcus aureus and Salmonella Typhimurium, using the agar diffusion method and by measuring the minimum inhibitory concentration, all the tested compounds showed antimicrobial activity against the three bacteria used. The modified aldehyde showed good values of a minimum inhibitory concentration of 0.03 mM and antimicrobial activity against Salmonella Typhimurium at 6.400 arbitrary units per milliliter (AU/mL). The identification of the antimicrobial properties of compounds derived from citronella suggests that the use of these substances to protect food against pathogenic bacteria deserves further exploration.     

Antibacterial activity and mechanism of bifidocin A against Listeria monocytogenes

Bifidocin A, produced by Bifidobacterium animalis BB04, is a novel bacteriocin with antimicrobial activity against a wide range of gram-positive and gram-negative foodborne bacteria. The objective of this study was to investigate the antibacterial activity and mechanism of action of bifidocin A against Listeria monocytogenes, one of the most susceptible bacteria to this bacteriocin. The minimum inhibitory concentration (MIC) of bifidocin A for L. monocytogenes 35152 was 0.029 mg/mL. Time-kill assays showed that bifidocin A effectively inhibited the growth of L. monocytogenes in a time-and concentration-dependent manner. The mechanism of action of bifidocin A was studied by analyzing its effects at a MIC on the cell morphology, intracellular organization, membrane permeability, membrane integrity, and membrane proton motive force (PMF) of L. monocytogenes. Scanning and transmission electron microscopy analyses showed that bifidocin A induced alterations in the morphology and intracellular organization of L. monocytogenes cells. Confocal laser scanning microscopy images showed that L. monocytogenes cells treated with bifidocin A took up propidium iodide. Bifidocin A treatment also induced the leakage of K⁺ and inorganic phosphate, the hydrolysis and release of ATP, and a collapse of the transmembrane electrical potential and pH gradient in L. monocytogenes cells. These results suggested that bifidocin A exerted its anti-Listeria monocytogenes effect through the dissipation of the cytoplasmic membrane PMF, increased membrane permeability, cell membrane pore formation, destruction of membrane integrity, and ultimately complete disintegration of the cells.     

Antimicrobial activity of polysaccharide films containing essential oils

Antimicrobial films were prepared by incorporating different concentrations of bergamot (BO), lemon (LO) and tea tree (TTO) essential oils (EO), into chitosan (CH) and hydroxypropylmethylcellulose (HPMC) films. Their antibacterial effectiveness against Listeria monocytogenes, Escherichia coli and Staphylococcus aureus was studied at 10 °C during a storage period of 12 days. HPMC-EO and CH-EO composite films present a significant antimicrobial activity against the three pathogens considered. The nature and amount of the essential oils (EO), the structure of the film and the possible interactions which exist between the polymers and active constituents of EO affected the antimicrobial activity of the films. In all film matrices, TTO exhibited the highest antimicrobial activity. A complete inhibition of microbial growth was observed for CH or HPMC-TTO films for E. coli, HPMC-TTO for L. monocytogenes and HPMC-BO for S. aureus.     

Investigation into the antibacterial behavior of suspensions of magnesium oxide nanoparticles in combination with nisin and heat against Escherichia coli and Staphylococcus aureus in milk

The antibacterial activities of magnesium oxide nanoparticles (MgO NP) alone or in combination with nisin against Escherichia coli and Staphylococcus aureus were investigated. Synergistic antibacterial effects existed and at lower levels of nisin when compared to when nisin was used alone. Also the antibacterial activities of MgO NP in combination with other antimicrobials (nisin and heat) against E. coli and S. aureus were investigated in milk. A synergistic effect of MgO NP in combination with nisin and heat was observed as well. Scanning electron microscopy was used to characterize the morphological changes of E. coli after antimicrobial treatments. It was revealed that MgO NP treatments in combination with nisin distort and damage the cell membrane, resulting in a leakage of intracellular contents and eventually the death of bacterial cells. This is the first report describing the antibacterial activity of MgO NPs and nisin in milk. It leads the way to development of treatment combinations which could result in a decrease in pasteurisation temperatures and the level of MgO NP required for pasteurising milk and maintaining pathogen control.     

Antibacterial properties of selenium nanoparticles and their toxicity to Caco-2 cells

The antibacterial properties of Se NPs were investigated against four foodborne pathogens (Escherichia coli O157:H7, Staphylococcus aureus, Salmonella, and Listeria monocytogenes). The cytotoxicity of Se NPs was also studied using the Caco-2 cell line. Se NPs at concentrations of 10 μg/mL or higher exhibited dose-dependent antimicrobial properties against S. aureus, but not on the other three pathogens. Se NPs also exhibited various degrees of toxicity on Caco-2 cells after 24 h of exposure. Cellular shrinkage and irregular shapes of treated bacterial cells indicated that the antimicrobial effects of Se NPs are bacteriostatic, not bactericidal. This is the first study to investigate the antimicrobial activity of Se NPs against important foodborne pathogens. The results of this study demonstrate that Se NPs can be used as an antimicrobial agent to inhibit the growth of S. aureus and can be potentially used for food safety applications.     

Antibacterial mechanism of bifidocin A,a novel broad-spectrum bacteriocin produced by Bifidobacterium animalis BB04

Bifidocin A, a novel broad-spectrum bacteriocin produced by Bifidobacterium animalis BB04, was isolated from the feces of a healthy centenarian. To understand the mechanism of the antibacterial action of bifidocin A against gram-negative bacteria, its effects at a minimum inhibitory concentration on cell morphology, intracellular organization, membrane permeability, membrane integrity, and membrane proton motive force (PMF) of Escherichia coli 1.90 were investigated. Scanning and transmission electron microscopy analyses showed that bifidocin A induced alterations in the morphology and intracellular organization of E. coli cells. The intracellular organization was more susceptible to changes induced by bifidocin A than the morphology. Bifidocin A treatment caused the leakage of K⁺ and inorganic phosphate, the release of ATP and UV-absorbing materials, and a collapse of the transmembrane electrical potential and pH gradient in E. coli cells. Confocal laser scanning microscopy images showed that E. coli cells treated with bifidocin A took up propidium iodide. These results suggested that the mechanism of action bifidocin A against E. coli involved dissipation of the PMF of the cytoplasmic membrane, an increase in membrane permeability, pore formation in the cell membrane, a change in membrane integrity, and complete cell disintegration.     

Bactericidal action of slightly acidic electrolyzed water against Escherichia coli and Staphylococcus aureus via multiple cell targets

This study combined plate counting method and fluorescent techniques (membrane integrity and potential, intracellular enzyme activity, and intracellular ROS level) to investigate the lethal and sublethal effects of slightly acidic electrolyzed water (SAEW) on Escherichia coli and Staphylococcus aureus. Also, the inactivation mechanism of SAEW was further explored through multiple cell targets (outer membrane and intracellular components). The results within 30 s SAEW treatment displayed 6.02 and 5.83 log reductions obtained for E. coli and S. aureus, respectively. The maximum sublethally injured cell proportions induced by SAEW exposure were 0.34 and 0.40 log₁₀ CFU/mL for E. coli and S. aureus, respectively. According to the data from experiments of various acting cellular sites by fluorescent techniques, SAEW damaged the microbial membrane integrity and membrane potential severely. Also, it posed inactivation effect on the activity of intracellular esterase enzymes. Therefore, SAEW showed disinfection behavior with multiple cellular targets, including both cell barriers and intracellular components. Furthermore, SAEW did not result in accumulation of reactive oxygen species (ROS) inside microbial cell, indicating SAEW conducted a ROS-independent behavior on microbial inactivation and the chemical oxidants (e.g., hypochlorous acid) played major role in microbial intracellular oxidation processing. The result in this study will help to further understand the disinfection mechanism underlying SAEW on microorganisms and make SAEW inactivation targets more explicit.     

Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi

The aim of this study was to investigate how mustard essential oil (EO) affected the cell membrane of Escherichia coli O157:H7 and Salmonella typhi. Intracellular pH and ATP concentration and the release of cell constituents were measured when mustard EO was in contact with E. coli and S. typhi at its minimal inhibitory concentration (MIC) and maximal tolerated concentration (MTC). The treatment with mustard EO affected the membrane integrity of bacteria and induced a decrease of the intracellular ATP concentration. Also, the extracellular ATP concentration increased and a reduction of the intracellular pH was observed in both bacteria. A significantly (P ? 0.05) higher release of cell constituent was observed when both bacteria cells were treated with mustard EO. Electronic microscopy observations showed that the cell membranes of both bacteria were apparently damaged by mustard EO. In conclusion, mustard EO affects the concentration of intracellular component, such as ATP in both bacteria and affects the pH suggesting that cytoplasmic membrane is involved in the antimicrobial action of mustard EO. Mustard EO can be used as an effective antimicrobial agent. We have demonstrated that mustard EO affected the cell membrane integrity, resulting in a loss of cell homeostasis.