Bacillus subtilis HJ18‐4 from Traditional Fermented Soybean Food Inhibits Bacillus cereus Growth and Toxin‐Related Genes

Bacillus subtilis HJ18‐4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad‐spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18‐4. Expression of B. cereus toxin–related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18‐4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18‐4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18‐4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus.     

Constituents,Oxidant-Antioxidant Profile,and Antimicrobial Capacity of the Essential Oil Obtained from Ferulago Sandrasica Peşmen and Quézel

Chemical constituents, total phenolic content, total oxidant status, total antioxidant status, lipid hydroperoxides, total free –SH levels, and antimicrobial activity of essential oil obtained from the Ferulago sandrasica (Umbelliferae) were investigated. The essential oil was obtained by hydrodistillation using a Clevenger-type apparatus. The chemical constituents were analyzed by gas chromatography‐mass spectrometry. The main components of the essential oil were ocimene (30.5%), carene-δ-3 (27.4%), and α-pinene (17.8). The antimicrobial activity was tested by a disc diffusion method against E. coli MC 400, E. coli ATCC 25922, E. coli 0157 H7, E. colaecea ATCC 23355, E. feacalis ATCC 19433, P. aeruginosa NRRL B-2679, S. aureus ATCC 25923, B. nischenoformis NRRL B-1001, S. aureus ATCC 33862, B. cereus NRRL B-3711, B. subtilis NRRL B-209, M. luteus NRRL B-1013, L. monocytogenes ATCC 7644, B. subtulis ATCC 6633.     

Hazel (Corylus avellana L.) leaves as source of antimicrobial and antioxidative compounds

Aqueous extracts of leaves of different hazel (Corylus avellana L.) cultivars (Cv. M. Bollwiller, Fertille de Coutard and Daviana), were analysed by reversed-phase HPLC/DAD for the definition of their phenolic composition. Antioxidant potential was assessed by the reducing power assay, and the scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and β-carotene linoleate model system. Their antimicrobial capacity was also tested against Gram positive (Bacillus cereus, Bacillus subtilis, Staphylococcus aureus) and Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae) and fungi (Candida albicans, Cryptococcus neoformans). Eight phenolic compounds were identified: 3-, 4- and 5-caffeoylquinic acids, caffeoyltartaric acid, p-coumaroyltartaric acid, myricetin-rhamnoside, quercetin 3-rhamnoside and kaempferol 3-rhamnoside. A p-coumaric acid, three myricetin and one quercetin derivatives were also detected. The hazel leaves extract presented high antioxidant activity in a concentration-dependent way, in general with similar behaviour of all cultivars. Gram positive bacteria revealed to be very sensitive to hazel leaf extract (MIC 0.1 mg/ml for B. cereus and S. aureus and 1 mg/ml for B. subtilis). However, Gram negative and the fungi displayed much lower sensitivity, being P. aeruginosa and C. albicans resistant at 100 mg/ml. Cv. M. Bollwiller exhibited the most potent antimicrobial activity.     

Antimicrobial and antioxidant activity of crude onion (Allium cepa,L.) extracts

Antioxidant and antimicrobial activity of the ethyl acetate and water subfractions of methanolic extracts of three Spanish onion varieties were assayed. Flavonoids were mainly present in ethyl acetate subfraction being 34.92 ± 0.75, 7.95 ± 0.16, 0.38 ± 0.01 μmol of rutin eq. g^?1 D.W. and its antioxidant capacity was 74.86 ± 1.77, 24.59 ± 0.67, 4.55 ± 0.44 μmol Trolox g^?1 D.W. of Grano de Oro, Fuentes de Ebro and Calçot de Valls varieties, respectively. The antimicrobial activity of flavonol standards and onion extracts was evaluated against some food spoiler microorganisms. Quercetin and kaempferol were inhibitory against gram positive bacteria such as Bacillus cereus, Staphylococcus aureus, Microcroccus luteus and Listeria monocytogenes. Gram negative bacteria Escherichia coli and Pseudomonas aeruginosa were less sensible to the antimicrobial effect of both flavonol standards and Candida albicans was totally resistant. Among the onion extracts tested only ethyl acetate subfraction showed antimicrobial inhibition.     

Isolation and characterization of an antimicrobial substance from Bacillus subtilis BY08 antagonistic to Bacillus cereus and Listeria monocytogenes

A wild-type bacteria producing an antimicrobial substance was isolated from Korean traditional fermented soybean paste, and identified as Bacillus subtilis. The antimicrobial substance purified by TLC, tentatively named UV254-B, displayed a specific antimicrobial activity against pathogenic Bacillus cereus and Listeria monocytogenes, without inhibiting the growth of soybean-fermenting Bacillus species. The antimicrobial substance was susceptible to proteinase K and lipase but resistant to esterase. Antimicrobial activity was observed over a wide range of pH from 3 to 11, with the maximum activity at pH 9, and thermal stability up to 80°C. The minimum inhibitory concentration of the antimicrobial substance was found to be 64 μg/mL for B. cereus and 128 μg/mL for L. monocytogenes. This antimicrobial substance has a putative molecular weight either at 1,133.6 or 1,700.5, which differs from that of other antimicrobial substances described for B. subtilis such as iturin, surfactin, fengycin, and subtilisin.     

In Vitro Antioxidant and Antibacterial Activities of Shallot and Scallion

ABSTRACT: In vitro antioxidant and antibacterial protection of shallot and scallion were examined. Water extract and oil of shallot and scallion significantly delayed lipid oxidation in multilamellar phosphatidylcholine liposomes and human RBC membranes (p < 0.05). Shallot oil at 5 and 10 mM showed marked antioxidant activity at 75 °C (p < 0.05). Shallot and scallion oils significantly inhibited the growth of 4 food‐borne bacteria, Salmonella typhimurium DT104, E.coli O157:H7, Listeria monocytogenes and Staphylococcus aureus, and 4 nosocomial bacteria, methicillin‐resistant Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii (p < 0.05). All observed antioxidant and antibacterial activities were dose‐dependent (p < 0.05). These results suggested the use of shallot and scallion oils in food systems could enhance lipid and microbial stability.     

Antimicrobia activity of garlic (Allium sativum L.) and holy basil (Ocimum sanctum L.) essential oils applied by liquid vs. vapour phases

Antimicrobial activity of garlic (Allium sativum L.) and holy basil (Ocimum sanctum L.) essential oils (GEO and HBEO, respectively) was evaluated in liquid vs. vapour phases. Diallyl disulphide and diallyl trisulphide in GEO while eugenol, caryophyllene and methyl eugenol in HBEO were predominant active compounds. Individually, GEO was highly effective in the vapour phase, showing the greatest antimicrobial activity (P ≤ 0.05) against Bacillus cereus and Staphylococcus aureus with complete inhibition, followed by Salmonella typhimurium and Pseudomonas fluorescens, but had no effect on Escherichia coli, Lactobacillus plantarum or Listeria monocytogenes. The combined GEO:HBEO (a 1:1 ratio) at 30 μL demonstrated inhibitive activity against all bacteria tested with complete inhibition against B. cereus and S. aureus. Minimum bactericidal concentration (MBC) of the combined oil was <0.4% v/v, except for P. fluorescens requiring a higher MBC (1.5%, v/v). This study demonstrated potential of the combined GEO:HBEO to be used for food preservation applications.     

Chemical composition,antioxidative capacity and interactive antimicrobial potency of Satureja khuzestanica Jamzad essential oil and antimicrobial agents against selected food‐related microorganisms

This study aimed to determine the chemical composition, antimicrobial and antioxidative activity of Satureja khuzestanica Jamzad essential oil. The oil was analysed by GC and GC/MS. Twenty‐eight constituents were identified. The oxygenated monoterpenes (78.22%) were the principal compound group. Among them, carvacrol (53.86%) and thymol (19.84%) were the most abundant constituents. The oil exhibited an acceptable antimicrobial activity against most of the tested microorganisms. The checkerboard method was applied to determine fractional inhibitory concentration indices (FICIs) to interpret the synergetic, additive, indifference or antagonistic interactions between essential oil and each of antimicrobials (lysozyme, ciprofloxacin, fluconazole and amphotericin B) against food‐related microorganisms. The synergetic phenomenon (FICI ≤ 0.5) was observed in majority of combinations with the exception of the essential oil and lysozyme. The oil exhibited good 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging activity (IC₅₀ = 28.71 μg mL^?1). Also, the oil had strong antioxidative activity in β‐carotene‐linoleic acid assay relative antioxidant activity (RAA% = 95.45). This study demonstrated that the essential oil has beneficial biological properties and its simultaneous application with standard antimicrobials against food‐related microorganisms result in reduction in inhibitory doses of the antimicrobials in vitro.     

Application of Green Technology for the Acquisition of Extracts of Araçá (Psidium grandifolium Mart. ex DC.) Using Supercritical CO2 and Pressurized Ethanol: Characterization and Analysis of Activity

Brazil is home to a wide variety of flora, including several lesser known species, such as araçá that were processed in this study using two green technologies consecutively: supercritical fluid extraction (SFE) for nonpolar fraction and pressurized liquid extraction. For polar fraction, the experiments followed a central composite design involving ethanol as solvent, with temperature and static time in each bath as independent variables. Both extracts were analyzed for antioxidant and antimicrobial activities. Total phenolics content (TPC) was determined for all ethanol extracts. In the araçá essential oil (AEO) obtained by SFE, the extraction yield was 2.33%. The three major compounds of AEO were α‐pinene (20.75%), p‐cymene (20.50%), and o‐cymene (20.05%). In ethanol extracts, the high yield (14.49%) was obtained at 74 °C/6 min and the major TPC (136.95 mg GAE/100 g) at 60 °C/9 min. Ethanol extracts presented good antioxidant activity (EC₅₀ = 6.37 mg/mL) at 74 °C/6 min. AEO was unable to reduce DPPH? concentration by 50%. Both extract types presented an inhibitory effect against Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus, while only the ethanol extracts presented effect against Listeria monocytogenes. This work had the aim to present the innovation of the use of a whole typical Brazilian fruit that gives rise to extracts with excellent properties for employment in both the pharmaceutical and food industries.     

Essential Oils Against Pathogen and Spoilage Microorganisms of Fruit Juices: Use of Versatile Antimicrobial Delivery Systems

Essential oils (EO) are increasingly used as natural antimicrobial compounds, however the effect of delivery system to enhance their antimicrobial activity has not been widely studied. Limonene (0 to 10 μL/mL) was added to microbial suspensions (～10⁵ CFU/mL) of selected foodborne pathogens (Listeria monocytogenes Scott A, Salmonella enterica Typhimurium, Escherichia coli and Staphylococcus aureus), and spoilage microorganisms (Lactobacillus plantarum, Saccharomyces cerevisiae, and Candida albicans). S. aureus was found to be the most sensitive foodborne pathogen while Salmonella enterica showed continued growth under all concentrations. Stable nanoemulsions and solid lipid nanoparticles (SLN) (d ～ 170 nm) were prepared using an alkane carrier oil (n‐tetradecane and n‐eicosane, respectively). Interfacial effects and homogenous distribution of limonene in nanoemulsions improved its (8 and 12 μL/mL) antimicrobial effect against S. aureus. Higher aqueous concentrations as a result of expulsion from SLN further enhanced the antimicrobial activity pronounced at higher limonene concentrations. Therefore, our findings confirm that the emulsion‐based delivery systems are able to effectively distribute limonene inside a microbial suspension to improve its antimicrobial activity.     

Antibacterial and antioxidant activities of the essential oil and methanol extracts of Bidens frondosa Linn

Antibacterial and antioxidant potential of essential oil, extract and its fractions of Bidens frondosa Linn were evaluated. Sixty‐one components representing 95.41% of the total oil were identified. The essential oil (7.5 μL disc^?1), methanol extract and its different organic subfractions (0.5 μg disc^?1) of B. frondosa displayed a great potential of antibacterial activity against Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes ATCC 19116, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella enteritidis KCTC 12021 and Enterobacter aerogenes KCTC 2190. Antioxidant activity was evaluated by using 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) assay. The free radical scavenging activity of ethyl acetate (EtOAc) fraction was superior to all other fractions (IC₅₀ = 11.96 μg mL^?1), which was higher than synthetic antioxidant butylated hydroxyanisole, (IC₅₀ = 18.27 μg mL^?1). Furthermore, the amount of total phenolic compounds was determined and its content in EtOAc fraction was the highest as compared to methanol extract or other fractions. The results indicate that the oil and extracts of B. frondosa could serve as an important bio‐resource of antimicrobial agents and antioxidants for using in the food industries.     

Composition, quality control and antimicrobial activity of the essential oil of cumin (Cuminum cyminum L.) seeds from Bulgaria that had been stored for up to 36 years

The essential oil of seeds of cumin (Cuminum cyminum L.) from Bulgaria stored for more than 35 years was analyzed by physicochemical methods, GC, GC‐MS and olfactometry and its antimicrobial activity tested using different strains of microorganisms. More than sixty constituents of this cumin oil could be identified as essential volatiles, responsible for the pleasant fresh, clean, spicy (typical cumin‐like) odour of a high quality product. Cumin aldehyde (36%), β‐pinene (19.3%), p‐cymene (18.4%) and γ‐terpinene (15.3%) were the principal compounds found. Antimicrobial testing showed high activity of the essential C. cyminum oil against the mold Aspergillus niger, the Gram (+) bacteria Bacillus subtilis and Staphylococcus epidermidis as well as the yeast Saccharomyces cerevisiae and Candida albicans.     

The effect of carbon dioxide on bacterial growth and on uptake processes in bacterial membrane vesicles

The effect of different concentrations of carbon dioxide on growth rates of Escherichia coli, Bacillus sublitis, Bacillus cereus and Pseudomonas aeruginosa was studied by means of a microculture system. For B. subtilis and P. aeruginosa, growth rate was linearly inhibited up to a concentration of 40% carbon dioxide. For E. coli and B. cereus, the inhibition of growth rate followed non-linear curves (concave downwards) in the whole concentration range (5–80%). Uptake of amino acids in E. coli and B. subtilis membrane vesicles was inhibited only half as much as growth, and uptake of glucose in E. coli membrane vesicles was unaffected even by 100% carbon dioxide. These experiments seem to indicate that the main mechanism of action for carbon dioxide is not associated with physical disturbance of the bacterial membrane.     

Applicability of bacteriocin‐producing Lactobacillus plantarum,Enterococcus faecium and Lactococcus lactis ssp. lactis as adjunct starter in Minas Frescal cheesemaking

The antimicrobial and technological characteristics of three bacteriocinogenic cultures used as adjunct starters in Minas Frescal cheese were investigated. The cheeses were manufactured with 1% commercial lactic starter and 0.5%Lactococcus lactis ssp. lactis ATCC 11454, Lactobacillus plantarum ALC 01 or Enterococcus faecium FAIR‐E 198. The cheeses were then artificially inoculated with Listeria monocytogenes Scott A, Staphylococcus aureus ATCC 27154 and Bacillus cereus K1‐B041 and stored for 21 days at 8°C. The results show that there was no significant difference in the counts of L. monocytogenes and S. aureus between the cheeses made with added bacteriocinogenic cultures and the control cheese. On the other hand, B. cereus exhibited susceptibility to Lb. plantarum ALC 01 and E. faecium FAIR‐E 198 from the seventh day of storage. However, these cultures increased the proteolysis of the Minas Frescal cheese.     

Assessment of Oligogalacturonide from Citrus Pectin as a Potential Antibacterial Agent against Foodborne Pathogens

Foodborne diseases are an important public health problem in the world. The bacterial resistance against presently used antibiotics is becoming a public health issue; hence, the discovery of new antimicrobial agents from natural sources attracts a lot of attention. Antibacterial activities of oligogalacturonide from commercial microbial pectic enzyme (CPE) treated citrus pectin, which exhibits antioxidant and antitumor activities, against 4 foodborne pathogens including Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Pseudomonas aeruginosa was assessed. Pectin hydrolysates from CPE hydrolysis exhibited antibacterial activities. However, no antibacterial activity of pectin was observed. Citrus oligogalacturonide from 24‐h hydrolysis exhibited bactericidal effect against all selected foodborne pathogens and displayed minimal inhibitory concentration at 37.5 μg/mL for P. aeruginosa, L. monocytogenes, and S. Typhimurium, and at 150.0 μg/mL for S. aureus.     

Antibacterial activity of chitosans with different degrees of deacetylation and viscosities

Antibacterial activities of six acid‐soluble [two degrees of deacetylation (DD) × three viscosities] and two water‐soluble chitosans (two DD with similar viscosities) were examined against eight gram‐negative (Pseudomonas fluorescens, Proteus vulgaris, Erwinia carotovora, Serratia marcescens, Escherichia coli, Vibrio parahaemolyticus, V. vulnificus, and Salmonella Typhimurium) and six gram‐positive bacteria (Listeria monocytogenes, Staphylococcus aureus, Bacillus subtilis, B. cereus, Lactobacillus curvatus, and L. plantarum). Antibacterial activities of chitosans differed depending on the chitosan types and bacteria tested. Water‐soluble chitosans inhibited bacterial growth by one to eight log cycles at 0.1% concentration; however, the effects were much lesser than those observed with 0.05% acid‐soluble chitosans. Minimum inhibitory concentration (MIC) values (0.03% to above 0.1%) of acid‐soluble chitosans were much lower than those (0.05% to above 0.8%) of water‐soluble chitosans. Based on MIC values, the acid‐soluble chitosan with 99% DD and lower viscosity (17.9 mPa s) was most effective in inhibiting bacteria growth among eight chitosans tested.     

Czech Ethanol‐Free Propolis Extract Displays Inhibitory Activity against a Broad Spectrum of Bacterial and Fungal Pathogens

Propolis acts primarily as a biocide against invasive bacteria and fungi in the hive, suggesting its potential for industrial applications. In food application, propolis is considered as a chemical preservative in meat products, extending shelf life of frozen meat and other food. The mechanism of action is still unclear due to the synergy of multiple compounds contained in propolis and due to parallel targeting of multiple pathways within each affected organism. Here, we examined the antimicrobial properties of dimethylsulfoxide (DMSO) Czech propolis extract. Until recently, DMSO was only rarely used in the propolis studies, although the other solvents tested (mostly ethanol) may significantly affect the observed inhibitory effects, notwithstanding the antimicrobial effects of ethanol itself. Here, we provide results of zone inhibition tests against Aspergillus fumigatus, Microsporum gypseum, Microsporum canis, Candida albicans, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. Although we determined inhibitory effects against all the microorganisms tested, the dose‐dependent response curves were not similar to each other. While inhibitory effects against C. albicans or S. aureus were strictly dose‐dependent, responses of M. gypseum and E. faecalis displayed plateau across the broad range of concentrations tested. Interestingly, response of E. coli revealed the double‐peak dose‐dependent curve, and responses of M. canis and L. monocytogenes decreased at the highest concentrations tested. Suggested is evaluation of DMSO propolis extracts in experimental treatment of human and veterinary infections, preferably in multitherapy with antibiotics.     

In vitro control of food-borne and food spoilage bacteria by essential oil and ethanol extracts of Lonicera japonica Thunb.

The antibacterial potential of essential oil from flowers and ethanolic leaf extracts of Lonicera japonica Thunb. was evaluated for controlling the growth of a range of food-borne pathogens. Thirty-nine compounds representing 92.34% of the total oil were identified, of which trans-nerolidol (16.31%), caryophyllene oxide (11.15%), linalool (8.61%), p-cymene (7.43%), hexadecanoic acid (6.39%), eugenol (6.13%), geraniol (5.01%), trans-linalool oxide (3.75%), globulol (2.34%), pentadecanoic acid (2.25%), veridiflorol (1.83%), benzyl alcohol (1.63%) and phenylethyl alcohol (1.25%) were the major compounds. The oil and extracts revealed a remarkable antibacterial effect against Listeria monocytogenes ATCC 19116, Bacillus subtilis ATCC 6633, B. cereus SCK 11, Staphylococcus aureus (ATCC 6538 and KCTC 1916), Salmonella enteritidis KCTC 12021, S. typhimurium KCTC 2515, Enterobacter aerogenes KCTC 2190 and Escherichia coli ATCC 8739. However, no effect was observed for Pseudomonas aeruginosa KCTC 2004 and E. coli O157:H7 ATCC 43888. Our findings demonstrate that the oil and extracts derived from L. japonica might be a potential source of preservatives for use in the food or pharmaceutical industries.     

Potential roles of essential oil and organic extracts of Zizyphus jujuba in inhibiting food-borne pathogens

This study was undertaken to examine the chemical compositions of essential oil and tested the efficacy of oil and organic extracts from seeds of Zizyphus jujuba against food-borne pathogens. The chemical compositions of the oil was analysed by Null. Twenty three compounds representing 91.59% of the total oil were identified. The oil (5 μl of 1:5 (v/v) dilution of oil with methanol) and extracts of hexane, chloroform, ethyl acetate and methanol (300 μg/disc) of Z. jujuba displayed a remarkable antibacterial activity against Staphylococcus aureus (ATCC 6538 and KCTC 1916), Listeria monocytogenes ATCC 19166, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa KCTC 2004, Salmonella typhimurium KCTC 2515 and Escherichia coli ATCC 8739. The scanning electron microscopic studies also demonstrated the effect of essential oil on the morphology of Staph. aureus ATCC 6538 at the MIC value, along with the potential effect on cell viabilities of the tested bacteria.