Formulation of food-grade microemulsions with glycerol monolaurate: effects of short-chain alcohols,polyols, salts and nonionic surfactants

The formulation of food-grade microemulsions with glycerol monolaurate (GML) for antimicrobial applications has been studied. The compositions included water, GML, short-chain alcohols (such as ethanol), polyols (propylene glycol and glycerol), salts (organic, such as sodium benzoate, sodium lactate and sodium propionate, and inorganic, such as NaCl) and several nonionic surfactants [such as ethoxylated sorbitan esters, sucrose esters (SEs)]. The phase behavior of these systems is discussed with respect to the influence of composition on the degree of oil solubilization in the aqueous phase. The oil solubilization was dramatically improved in the presence of the short-chain alcohols and polyols, organic salts contributed to the improvement of oil solubilization as hydrotropes. Tween 20 being the most hydrophilic surfactant in Tweens solubilized the maximum oil, but in the presence of SEs it is hard to form oil-in-water microemulsions. Viscosity measurements along selected dilution lines indicate that at a certain composition the system inverts from a water-in-oil to an oil-in-water microemulsion.     

单月桂酸甘油酯微乳液抗菌性研究

单月桂酸甘油酯(GML)作为一种食品级乳化剂也具有一定抗菌作用。本研究将GML与其它配合物复合,使其抗菌性得到不同程度提高,通过Box–Behnken Design–响应面优化分析设计,得出抗菌性最佳GML乳化液配方为:以浓度为50μg/mL 10 ml GML为基质,山梨酸钾加入量0.050 g、吐温–80加入量0.022 g、SE–15加入量0.030 gS、SL加入量0.025 g,在此乳化液中加入盐、乙醇和水制成GML微乳液。实验证明,其具有优良抗菌效果,应用到馒头制作中也得到良好防霉保鲜效果,为GML在食品中应用提供新的思路。     

Antimicrobial activity of a food-grade fully dilutable microemulsion against Escherichia coli and Staphylococcus aureus

Microemulsions are colloidal nanodispersions of oil and water stabilized by an interfacial film of surfactant molecules, typically in conjunction with a cosurfactant. There is a limited number of reports in the literature on microemulsion use for antimicrobial purposes. The physicochemical characterization of a food-grade fully dilutable microemulsion system with glycerol monolaurate (GML) as oil, organic acids as cosurfactant, Tween 80 as surfactant, and the antimicrobial activities against Escherichia coli and Staphylococcus aureus have been studied in this paper. The influence of organic acids on oil solubilization was clearly reflected in the phase behavior of these systems. Propionic acid demonstrated the greatest capability to improve the oil solubilization among the tested linear and nonlinear chain organic acids and contributed to the formation of U-type microemulsion systems. One microemulsion formulation with an average particle size of 8 nm was selected, the composition is GML/propionic acid/Tween 80/water = 3:9:8:12. The kinetics of killing experiments demonstrated that the undiluted microemulsion caused a complete loss of viability of E. coli or S. aureus cells in 1 min and still had effective bactericidal effects even when diluted, more than 99% viable E. coli cells were killed within 15 min and a complete loss of viability was achieved at 45 min while more than 99% viable S. aureus cells were killed within 30 min and a complete loss of viability was achieved at 60 min in the presence of the 10-fold diluted microemulsion. The fast killing kinetics of the ten-fold serial dilutions of microemulsions were in good agreement with the mode of action studies, indicating that the interaction between the antimicrobial microemulsions and bacterial membranes significantly decreased the bacterial cell surface hydrophobicity and induced the quick release of 260 nm absorbing materials. This work suggests the potential use of food-grade fully dilutable microemulsions for antimicrobial purposes in beverages or seafood products.     

In vitro antimicrobial effects and mechanism of action of selected plant essential oil combinations against four food-related microorganisms

The aim of this study was to evaluate the antimicrobial efficacy of selected plant essential oil (EO) combinations against four food-related microorganisms. Ten EOs were initially screened against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Saccharomyces cerevisiae using agar disk diffusion and broth dilution methods. The highest efficacy against all the tested strains was shown when testing the oregano EO. EOs of basil and bergamot were active against the Gram-positive bacteria (S. aureus and B. subtilis), while perilla EO strongly inhibited the growth of yeast (S. cerevisiae). The chemical components of selected EOs were also analyzed by GC/MS. Phenols and terpenes were the major antimicrobial compounds in oregano and basil EOs. The dominant active components of bergamot EO were alcohols, esters and terpenes. For perilla EO, the major active constituents were mainly ketones. The checkerboard method was then used to investigate the antimicrobial efficacy of EO combinations by means of the fractional inhibitory concentration index (FICI). Based on an overall consideration of antimicrobial activity, organoleptic impact and cost, four EO combinations were selected and their MIC values were listed as follows: oregano–basil (0.313–0.313 μl/ml) for E. coli, basil–bergamot (0.313–0.156 μl/ml) for S. aureus, oregano–bergamot (0.313–0.313 μl/ml) for B. subtilis and oregano–perilla (0.313–0.156 μl/ml) for S. cerevisiae. Furthermore, the mechanisms of the antimicrobial action of EO combinations to the tested organisms were studied by the electronic microscopy observations of the cells and the measurement of the release of cell constituents. The electron micrographs of damaged cells and the significant increase of the cell constituents' release demonstrated that all EO combinations affected the cell membrane integrity.     

Preparation Methods for Monodispersed Garlic Oil Microspheres in Water Using the Microemulsion Technique and Their Potential as Antimicrobials

Garlic oil is considered as a natural broad‐spectrum antibiotic because of its well‐known antimicrobial activity. However, the characteristics of easy volatility and poor aqueous solubility limit the application of garlic oil in industry. The purpose of the present work is to develop and evaluate an oil‐free microemulsion by loading garlic oil in microemulsion system. Microemulsions were prepared with ethoxylated hydrogenated castor (Cremophor RH40) as surfactant, n‐butanol (or ethanol) as cosurfactant, oleic acid‐containing garlic oil as oil phase, and ultrapure water as water phase. The effects of the ratio of surfactant to cosurfactant and different oil concentration on the area of oil‐in‐water (O/W) microemulsion region in pseudoternary phase diagrams were investigated. The particle size and garlic oil encapsulation efficiency of the formed microemulsions with different formulations were also investigated. In addition, the antimicrobial activity in vitro against Escherichia coli and Staphylococcus aureus was assessed. The experimental results show that a stable microemulsion region can be obtained when the mass ratio of surfactant to cosurfactant is, respectively, 1:1, 2:1, and 3:1. Especially, when the mixture surfactants of RH40/n‐butanol 2/1 (w/w) is used in the microemulsion formulation, the area of O/W microemulsion region is 0.089 with the particle size 13.29 to 13.85 nm and garlic oil encapsulation efficiency 99.5%. The prepared microemulsion solution exhibits remarkable antibacterial activity against S. aureus.     

Antimicrobial activity of greater galangal [Alpinia galanga (Linn.) Swartz.] flowers

Using the agar disc diffusion method, the potential antimicrobial activity of edible galangal [Alpinia galanga (Linn.) Swartz.] flower against Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, and Shigella, and the effects of different drying methods and solvent types on the flowers’ antimicrobial activity were investigated. Oven-dried ethanol (OD Ethanol) extract from galangal flower was the most effective against S. aureus with inhibition zone of about 26–31 mm and the minimum inhibitory concentration (MIC) ranging from 0.352–0.547 mg/mL. No antimicrobial activity was observed on E. coli O157:H7 and Salmonella. Overall antimicrobial activity of oven-dried samples extracted with ethanol (OD Ethanol) was the highest with inhibition zone of 8.94 mm and MIC of 1.457 mg/mL. In contrast, freeze-dried samples extracted with ethanol (FD Ethanol) exhibited the lowest overall antimicrobial activity (7.05 mm and 2.470 mg/mL). This is the first report describing antimicrobial activity of galangal flowers against Grampositive S. aureus.     

Antioxidant and antimicrobial activities of Polygonum cognatum Meissn extracts

The antioxidant activities, reducing powers, 2,2‐diphenyl‐l‐picrylhydrazyl (DPPH) radical‐scavenging activities, total phenolic compound contents and antimicrobial activities of ether, ethanol and hot water extracts of Polygonum cognatum Meissn were studied in vitro. The highest antioxidant activity was found in the water extract. However, there were no statistically significant differences among 15 µg ml^?1 extract‐containing samples in linoleic acid emulsion (0.02 M , pH 7.0) during 120 h of incubation (P > 0.05). The reducing power of the water extract was the highest, but its reducing power was markedly lower than that of ascorbic acid. The highest DPPH radical‐scavenging activity was found in the water extract, with 50% DPPH radical scavenging at a concentration of 100 µg ml^?1 dried water extract, while at the same concentration of dried ethanol extract the value was 12%. Surprisingly, no DPPH radical‐scavenging activity was observed in the ether extract. The concentrations of phenolic compounds found were 0.48, 0.50 and 0.01 µg ml^?1 gallic acid equivalent in 10 µg ml^?1 water, ethanol and ether extracts respectively. The ether and ethanol extracts showed antimicrobial activity against Staphylococcus aureus and Bacillus subtilis. The water extract did not show antimicrobial activity against the studied micro‐organisms. © 2002 Society of Chemical Industry     

Phytochemical analysis,in vitro antioxidant and antimicrobial activities of male flower of Juglans regia L.

The male flower of Juglans regia L., were investigated for its in vitro antioxidant activity, antimicrobial activity, and chemical constituents. The antioxidant activity showed that the methanol extract of J. regia male flower (MEJR) had highest scavenging potential than the other solvents (ethanolic = EEJR and aqueous = AEJR). The antimicrobial activity showed that Staphylococcus aureus and Escherichia coli were the most sensitive organisms and significant activity was also recorded against both the fungal strains tested, with highest activity against Candida albicans. Totally, 26 constituents were identified by high-resolution-liquid chromatography-mass spectrometry analyses from which seven compounds were identified first time from the extract.     

A new antimicrobial substance produced by Staphylococcus pasteuri isolated from vegetables

Staphylococcus aureus is important food-borne pathogen in agricultural products consumed in a fresh state. The antimicrobial activities of staphylococci isolated from vegetables in Korea against S. aureus were investigated. Coagulase-negative staphylococci (CNS) isolates (12) with antimicrobial activities against S. aureus were identified as S. xylosus, pasteuri, and epidermidis. CNS isolates exhibited antimicrobial activities against Gram-positive bacteria. Inactivation of S. pasteuri antimicrobial activity by proteases suggested that the substances evaluated were antimicrobial peptides or bacteriocins. None of the S. pasteuri isolates possessed homologous DNA fragments known to be bacteriocin genes, except for nukacin. The nukM gene that encodes a post-translational modification enzyme in the nukacin gene cluster was not detected in S. pasteuri isolates. S. pasteuri probably produces one or more new antimicrobial substances.     

Enhancement of antimicrobial and antimutagenic activities of Korean barberry (Berberis koreana Palib.) by the combined process of high‐pressure extraction with probiotic fermentation

BACKGROUND: To evaluate the combined effects of high pressure extraction (HPE) and probiotic fermentation on the antimicrobial and antimutagenic activities, Berberis koreana was subjected to 500 MPa for 30 min and then fermented with Bifidobacterium longum B6 (HPE‐BLF) and Lactobacillus paracasei (HPE‐LPF) at 37 °C for 6 days. RESULTS: The phenol content was significantly increased to 228 mg GAE g^?1 by the HPE compared to the conventional extraction (CE, 188 mg GAE g^?1). The HPE‐BLF and HPE‐LPF showed the highest antimicrobial activity (MIC < 4 mg mL^?1) against β‐lactam antibiotic sensitive and resistant Staphylococcus aureus. No significant mutagenic effect was observed for CE, HPE, HPE‐BLF, and HPE‐LPF extracts. The highest antimutagenic activities against frame‐shift mutant Salmonella typhimurium were observed at the HPE‐LPF (82%), followed by the HPE‐BLF (77%). CONCLUSION: The combined HPE and fermentation process could be used as an alternative extraction method for improving the extraction efficacy of medicinal plants. The results will provide pharmaceutically useful information and potential direction for finding new drug sources from medicinal plants. Copyright © 2010 Society of Chemical Industry     

Inhibition of Staphylococcus aureus by Phenolic Phytochemicals of Selected Clonal Herbs Species of Lamiaceae Family and Likely Mode of Action through Proline Oxidation

Staphylococcus aureus, a Gram-positive organism, is responsible for numerous infections ranging in severity from skin and soft tissue infections to endocarditis and septic arthritis. Further, Staphylococcal food poisoning (SFP) caused by enterotoxigenic staphylococci is one of the main food-borne diseases. In this study, several single-seed origin clonal lines of herbs from Lamiaceae family were screened for antimicrobial applications against S. aureus. These clonal lines have consistent total phenolic content, phenolic profile and antioxidant activity as opposed to heterogenous phenolic profiles of typical herbal extracts. The clonal extracts of specific herb species were evaluated for anti- S. aureus activity. Water extracts of clonal lemon balm had the highest anti- S. aureus activity followed by clonal extracts of rosemary RoLA, sage, oregano, rosemary Ro-6, rosemary RoK-2 and chocolate mint. The anti- S. aureus activity of the water extracts correlated with gallic acid and caffeic acid content, which in pure form showed high anti- S. aureus activity. Based on the rationale that gallic acid and caffeic acid were likely proline analogs affecting proline oxidation, the countering effect of proline to overcome their anti- S. aureus activity was evaluated. Results indicated proline can overcome the inhibitory effects of gallic acid and caffeic acid. These results point to the likely role of phenolic oxidation via proline dehydrogenase, which could be an important target for inhibition by phenolic phytochemicals behaving as proline analog mimics. This mode of regulation by phenolics at the bacterial proline dehydrogenase in the plasma membrane may be an important antimicrobial defense in plants and has consequences for disruption of critical energy metabolism of invading bacterial pathogen. This rationale could be used to design new antimicrobial strategies against S. aureus and other relevant bacterial pathogens.     

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.     

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.     

Bactericidal Activities of Health‐Promoting,Food‐Derived Powders Against the Foodborne Pathogens Escherichia coli,Listeria monocytogenes,Salmonella enterica,and Staphylococcus aureus

We evaluated the relative bactericidal activities (BA₅₀) of 10 presumed health‐promoting food‐based powders (nutraceuticals) and, for comparison, selected known components against the following foodborne pathogens: Escherichia coli O157:H7, Salmonella enterica, Listeria monocytogenes, and Staphylococcus aureus. The relative activities were evaluated using quantitative bactericidal activity [(BA₅₀ value, defined as the percentage of the sample in the assay mixture that resulted in a 50% decrease in colony forming units]. The BA₅₀ values were determined by fitting the data to a sigmoidal curve by regression analysis using concentration–antimicrobial response data. Antimicrobial activity is indicated by a low BA₅₀ value; meaning less material is needed to kill 50% of the bacteria. Olive pomace, olive juice powder, and oregano leaves were active against all 4 pathogens, suggesting that they behave as broad‐spectrum antimicrobials. All powders exhibited strong antimicrobial activity against S. aureus. The following powders showed exceptionally high activity against S. aureus (as indicated by the low BA₅₀ values shown in parentheses): apple skin extract (0.002%); olive pomace (0.008%); and grape seed extract (0.016%). Listeria bacteria were also highly susceptible to apple skin extract (0.007%). The most active substances provide candidates for the evaluation of antimicrobial effectiveness in human food and animal feed. Practical Application : Plant‐derived health‐promoting food supplements, high in bioactive compounds, are candidates for use as antimicrobials in food.     

Antioxidant and antimicrobial activities of Smilax china L. leaf extracts

Antioxidant and antimicrobial activities of Smilax china L. leaf extracts obtained with methanol, ethanol, acetone, and water were investigated. Antioxidant activity was evaluated by determining the DPPH radical scavenging activity, ABTS radical scavenging activities, total phenol content (TPC), and reducing power (RP). The highest DPPH, ABTS radical scavenging activity, and RP were found in the ethanol extract, which also showed the highest TPC (105.81±0.48 μg gallic acid equivalents/mL). The antimicrobial activity of all the extracts against foodborne microorganisms was determined by paper disc method. All the extracts inhibited the growth of Listeria monocytogenes, Staphylococcus aureus, and Salmonella Typhimurium, however, no antimicrobial activity was observed against Escherichia coli O157:H7. The results indicated that Smilax china L. possessed antioxidant and antimicrobial substances, and suggested that the ethanol extract can be applied into food and cosmetic industry.     

Screening for Alternative Antibiotics: An Investigation into the Antimicrobial Activities of Medicinal Food Plants of Mauritius

ABSTRACT: The present study was designed to evaluate the antimicrobial activities of 2 endemic medicinal plants; Faujasiopsis flexuosa (Asteraceae) (FF) and Pittosporum senacia (Pittosporaceae) (PS) and 2 exotic medicinal plants, Momordica charantia (Cucurbitaceae) (MC) and Ocimum tenuiflorum (Lamiaceae) (OT) that forms part of local pharmacopoeia of Mauritius and correlate any observed activity with its phytochemical profile. Aqueous and organic fractions of the leaves, fruits, and seeds of these plants were subjected to antimicrobial testing by the disc diffusion method against 8 clinical isolates of bacteria and 2 strains of fungus. It was found that MC, OT, and FF possessed antimicrobial properties against the test organisms. The MIC for MC ranged from 0.5 to 9 mg/mL and that of FF from 2 to 10 mg/mL and the lowest MIC value (0.5 mg/mL) was recorded for the unripe fruits of MC against E. coli. On the other hand, higher concentration of the unripe MC fruit extract of 9 mg/mL was needed to be effective against a resistant strain of Staphylococcus aureus (MRSA). The antimicrobial effect against MRSA was lost upon ripening of the fruits. The methanolic extract of both MC and FF showed highest MIC values compared to the corresponding aqueous extract, which indicates the low efficacy and the need of higher doses of the plant extract. Phytochemical screening of the plants showed the presence of at least tannins, phenols, flavonoids, and alkaloids, which are known antimicrobial phyto-compounds. In conclusion, the observed antimicrobial properties would tend to further validate the medicinal properties of these commonly used endemic medicinal and food plants of Mauritius.     

Influence of process variables on antimicrobial properties of cotton knitted fabrics

ABSTRACT

This work discusses the influence of process variables on antimicrobial properties of cotton single jersey knitted fabrics. The antimicrobial process variables are optimized based on the Box-Benken experimental design using design-expert software 8.0 to predict the optimum process parameter for achieving the required level of antimicrobial properties. The effects of process parameters such as yarn count, loop length, and finishing % of the zone of inhibition (ZOI) against the Escherichia coli and Staphylococcus aureus microbes on cotton knitted fabric have been carried out. Result shows that except yarn count other parameters are not significant toward the ZOI against microbes.     

Hurdle Effect of Antimicrobial Activity Achieved by Time Differential Releasing of Nisin and Chitosan Hydrolysates from Bacterial Cellulose

We investigated the combined antimicrobial effect of nisin and chitosan hydrolysates (CHs) by regulating the antimicrobial reaction order of substances due to differential releasing rate from hydroxypropylmethylcellulose‐modified bacterial cellulose (HBC). The minimum inhibitory concentration of nisin against Staphylococcus aureus and that of CHs against Escherichia coli were 6 IU and 200 μg/mL, respectively. Hurdle and additive effects in antimicrobial tests were observed when nisin was used 6 h before CH treatment against S. aureus; similar effects were observed when CH was used before nisin treatment against E. coli. Simultaneously combined treatment of nisin and CHs exhibited the low antimicrobial effect. HBC was then selected as the carrier for the controlled release of nisin and CHs. A 90% inhibition in the growth of S. aureus and E. coli was achieved when 30 IU‐nisin‐containing HBC and 62.5 μg/mL‐CH‐containing HBC were used simultaneously. The controlled release of nisin and CHs by using HBC minimized the interaction between nisin and CHs as well as increased the number of microbial targets.     

Development of antimicrobial gummy candies with addition of bovine colostrum,essential oils and probiotics

The aim of this study was to develop antimicrobial properties of gummy candies based on bovine colostrum (BC), essential oils (EOs), lactic acid bacteria (LAB) strains and their combinations. In addition, the heteropolysaccharide (agar), as a multifunctional polymer, was used for the antimicrobial candies preparation. The antimicrobial activities of BC, EOs (C. reticulata L., Eugenia caryophyllata, C. paradisi L., Thymus vulgaris), LAB strains (Lactobacillus plantarum LUHS135 and Lactobacillus paracasei LUHS244) and their combinations against pathogenic bacteria (Pseudomonas aeruginosa, Proteus mirabilis, Escherichia coli, Salmonella enterica, Staphylococcus aureus, Enterococcus faecalis, Streptococcus mutans) were investigated. The highest antimicrobial activities were demonstrated by Thymus vulgaris and Eugenia caryophyllata EOs and their emulsions (12%), and the best formulation of components for antimicrobial gummy candies production would incorporate the BC fermented with L. paracasei LUHS244 in combination with Thymus vulgaris or Eugenia caryophyllata EOs, which inhibited growth of all the tested pathogenic microorganisms (except Pseudomonas aeruginosa). Gummy candies formula consisting of the fermented BC (up to 3%) and thyme EO (up to 0.2%) with mandarin or grapefruit EOs (up to 0.2%) for taste‐masking, allowed obtaining good texture and high overall acceptability products containing desirable antimicrobials, thus antimicrobial gummy candies could be consumer preferred form of nutraceuticals.     

Characterization of antimicrobial polylactic acid based films

Olive leaf extract (OLE) (Olea europaea L.), which has antimicrobial effect on many food pathogens, was incorporated as antimicrobial agent into polylactic acid (PLA) films. Antimicrobial activities of films were tested against Staphylococcus aureus. Increasing amount of the OLE in the film discs from 0.9 mg to 5.4 mg caused a significant increase in inhibitory zones from 9.10 mm to 16.20 mm, respectively. Moreover, incorporation of OLE and/or increasing the amount in the film formulation significantly enhanced the water vapor permeability (WVP). The water solubility and the degradation rates of films increased up to 19.3% and 22.4%, respectively. Thus, OLE incorporated PLA films have a prospectively potential in antimicrobial food packaging to reduce post-process growth of S. aureus with improved properties.