Chemotaxonomical analysis of the essential oil aroma compounds of four different<Emphasis Type="Italic"> Ocimum</Emphasis> species from southern India

The essential oil aroma compounds of four different Ocimum species ( O. americanum, O. basilicum, O. gratissimum and O. sanctum) from southern India were analysed by solid phase microextraction (SPME)/gas chromatography (GC)/flame ionization detection (FID), SPME/GC/mass spectrometry (MS) and olfactoric evaluations. The essential oil of the whole plant of O. americanum is rich in ( E)-methyl cinnamate (72.05%), ( Z)-methyl cinnamate (9.11%), camphor (5.95%) and ß-selinene (3.43%); the essential leaf oil of O. basilicum contains ( E)-methyl cinnamate (34.49%), linalool (28.44%), camphor (13.08%), ( Z)-methyl cinnamate (6.90%) and geraniol (3.84%), whereas the essential leaf oil of O. gratissimum comprises eugenol (63.36%), ( Z)-ß-ocimene (9.11%), germacrene D (8.84%) and ß-caryophyllene (3.89%). Finally, the essential leaf oil of O. sanctum shows methyl eugenol (56.18%), ß-caryophyllene (16.60%) and germacrene D (5.10%) as main constituents. Therefore, the following chemotypes can be attributed to the analysed Ocimum samples: O. americanum, methyl cinnamate-type; O. basilicum, methyl cinnamate/linalool-type; O. gratissimum, eugenol-type; and O. sanctum, methyl eugenol-type. The character impact compounds of the essential oils of the four investigated Ocimum samples, as well as a discussion of their possible use in food products, is also given.     

Chemical composition and antimicrobial activity of the essential oils of four Ocimum species growing in Tanzania

As part of ongoing research on Tanzanian plants used as edibles or spices, six samples of essential oils from four Ocimum species (O. basilicum, O. kilimandscharicum, O. lamiifolium, O. suave) were analyzed by GC and GC–MS. Eighty-one compounds, corresponding to 81.1–98.2% of the chemical components of the oils, were identified. Major compounds were either phenyl propane derivatives or terpenoids, including methyl eugenol, 1,8-cineole, camphor, bornyl acetate, germacrene-D, E-myroxide, germacrene-B, caryophylene oxide and p-cymene. The oils were also evaluated for antimicrobial activity against eight bacterial strains and three fungi. The oil of O. suave (B) showed the strongest antibacterial activity; O. suave (A), O. kilimandscharicum and, O. lamiifolium were moderately active, while O. basilicum oil was weakly active. However, none of the oils was active against the fungi species. The study has shown that, Ocimum oils could potentially be used as anti-infective agents.     

In vitro application of selected essential oils and their major components in controlling fungal pathogens of crown rot in Embul banana (Musa acuminata– AAB)

The necessity for pesticide‐free fresh produce have prompted investigating the effect of selected essential oils and their major components on inhibition of conidial germination, appressoria formation and membrane permeability changes of the pathogens responsible for crown rot of banana. Eugenol, which is the major component of the essential oil of Ocimum basilicum, was the most effective chemical component in inhibiting conidial germination of Colletotrichum musae and Fusarium proliferatumin vitro while Cymbopogon citratus oil was the least effective. Both O. basilicum and C. citratus oils and their major components (Eugenol, citral a + b) inhibited appressoria formation by C. musae and changed the selective permeability of conidial membranes. Ocimum basilicum oil, eugenol and citral a + b could be satisfactorily used for inhibition of conidial germination and disruption of conidial activity of banana pathogens.     

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 aromatic compounds and biological activities of essential oils from Tunisian aromatic plants

The aromatic compounds and biological activities of essential oils from six Tunisian aromatic plants including Artemisia herba-alba, Rosmarinus officinalis, Thymus capitatus, Mentha piperita, Ocimum basilicum and Artemisia absinthium were investigated. Hydro-distillation was used to extract essential oil from these plants. The identification of compounds from essential oils was performed using GC–MS analysis. Camphor (28.47%) was the major compound of A. absinthium essential oil. High contents of verbenone (20.99%) and camphor (19.72%) were found in R. officinalis. In the case of T. capitatus, carvacrol (81.09%), gamma terpinene (6.61%) and caryophyllene (4.87%) were identified as the major compounds. While eugenol (24.69%), linalool (18.00%) were characteristic compounds of O. basilicum essential oil, camphor (39.10%) and farnesol (14.25%) together with bornyl acetate (12.31%) were the main constituents of A. absinthium. These oils were also subjected to a screening for their antioxidant activity and essential oil from A. absinthium showed a greater antioxidant activity (IC₅₀?=?0.0063 mg/mL) compared to the standard Vitamin E (IC₅₀?=?0.019 mg/mL). The antibacterial activities of the oils against seven pathogenic strains, Pseudomonas aeruginosa, Bacillus cereus, Salmonella, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Micrococcus luteus, were tested. The highest and broadest activity was shown by M. piperita; however, Ocimum basilicum was inactive against all strains. Essential oils were also evaluated for antidiabetic and anti-acetylcholinesterase activities. The IC₅₀ values of A. herba-alba and O. basilicum against α-amylase were respectively 17.76 and 16.32 µg/mL suggesting a powerful anti-diabetic effect comparable to that of acarbose (IC₅₀?=?14.88 µg/mL). R. officinalis, M. piperita and A. absinthium exhibited an interesting acetylcholinesterase inhibition with IC₅₀ equal to 22, 24 and 58 µg/mL respectively.     

Antimicrobial activity in the vapour phase of a combination of cinnamon and clove essential oils

The antimicrobial activity of the vapour generated by a combination of cinnamon and clove essential oils against the growth of four Gram-negative (Escherichia coli, Yersinia enterocolitica, Pseudomonas aeruoginosa and Salmonella choleraesuis) and four Gram-positive bacteria (Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus and Enterococcus faecalis) was assessed by means of the fractional inhibitory concentration index (FIC) of the mixture. The presence of synergism or antagonism effects depended on the reference parameter used to estimate such an index. If the minimal inhibitory concentrations were applied, the vapours of the combination of essential oils exerted an antagonistic effect on the growth of E. coli, while they wielded a synergistic effect for the inhibition of L. monocytogenes, B. cereus and Y. enterocolitica when the concentrations of maximal inhibition were used. This fact revealed a clear concentration-dependent interaction.     

Partial Characterization of Nine Bacteriocins Produced by Lactic Acid Bacteria Isolated from Cold-Smoked Salmon with Activity against Listeria monocytogenes

Nine LAB bacteriocin-producers, isolated from vacuum-packaged cold-smoked salmon (CSS), were phenotypically and genotypically identified as Lactobacillus curvatus, Lactobacillus delbrueckii, Lactobacillus fermentum, Enterococcus faecium, and Pediococcus acidilactici. Their bacteriocins were partially characterized. The antimicrobial spectrum was determined against Listeria monocytogenes, E. faecalis, E. faecium, and Staphylococcus aureus. The molecular size of bacteriocins ranged from 2.8 to 4.5 kDa. They were inactivated by treatment with proteolytic enzymes but not by lipolytic or glycolytic enzymes. Maximal activity against L. monocytogenes ranged between 800 and 10000 AU/mL at pH 6.5. Most of the bacteriocins maintained full activity in a pH range of 2.0 to 8.0 but were partially or completely inactivated at pH 10.0. After heating at 60°C and 100°C, only two bacteriocins from Lb. curvatus strains partially lost activity. All bacteriocins showed a narrow spectrum of activity and a high anti-listerial activity, which is characteristic of the class IIa bacteriocins. Isolated bacteriocin-producing LAB could be used successfully in the bio-preservation of CSS and development of new potential bio-preservatives for CSS active against L. monocytogenes.     

Bacterial diversity of Darfiyeh, a Lebanese artisanal raw goat's milk cheese

In order to contribute to the preservation of the Lebanese dairy heritage, the aim of this study was to characterize the Darfiyeh cheese, a traditional variety made from raw goat's milk and ripened in goat's skin. Three independent batches of Darfiyeh production were analyzed after 20, 40 and 60 days of ripening. Mesophilic lactobacilli, thermophilic coccal-shaped lactic acid bacteria (LAB) and thermophilic lactobacilli were enumerated. In order to explore the Darfiyeh natural ecosystem, a combination of phenotypical and molecular approaches was applied. The latter included Polymerase Chain Reaction-temporal temperature gel electrophoresis (PCR-TTGE), classical PCR and quantitative PCR. These methods revealed the presence of Streptococcus thermophilus, Enterococcus faecium, Enterococcus durans, Enterococcus faecalis, Enterococcus malodoratus, group D Streptococcus sp., Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris, Lactobacillus plantarum, Lactobacillus curvatus, Staphylococcus haemolyticus, Escherichia coli, Clostridium sp./Eubacterium tenue. Real-time PCR enabled quantification of E. faecium, with a detection of 10⁷–10⁹ cfu g⁻¹ of product. The present molecular approaches combined with phenotypic method allowed describing the complex natural ecosystem of Darfiyeh, giving useful information for the preservation of Lebanese artisanal dairy products.     

Incorporation of Essential Oils and Nanoparticles in Pullulan Films to Control Foodborne Pathogens on Meat and Poultry Products

The incorporation of essential oils and nanotechnology into edible films has the potential to improve the microbiological safety of foods. The aim of this study was to evaluate the effectiveness of pullulan films containing essential oils and nanoparticles against 4 foodborne pathogens. Initial experiments using plate overlay assays demonstrated that 2% oregano essential oil was active against Staphylococcus aureus and Salmonella Typhimurium, whereas Listeria monocytogenes and Escherichia coli O157:H7 were not inhibited. Two percent rosemary essential oil was active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 1%. Zinc oxide nanoparticles at 110 nm were active against S. aureus, L. monocytogenes, E. coli O157:H7, and S. Typhimurium, when compared with 100 or 130 nm. Conversely, 100 nm silver (Ag) nanoparticles were more active against S. aureus than L. monocytogenes. Using the results from these experiments, the compounds exhibiting the greatest activity were incorporated into pullulan films and found to inhibit all or some of the 4 pathogens in plate overlay assays. In challenge studies, pullulan films containing the compounds effectively inhibited the pathogens associated with vacuum packaged meat and poultry products stored at 4 °C for up to 3 wk, as compared to control films. Additionally, the structure and cross‐section of the films were evaluated using electron microscopy. The results from this study demonstrate that edible films made from pullulan and incorporated with essential oils or nanoparticles may improve the safety of refrigerated, fresh or further processed meat and poultry products.     

Safety Assessment and Probiotic Evaluation of Enterococcus Faecium YF5 Isolated from Sourdough

Enterococcus faecium YF5, a strain previously isolated from sourdough, was assessed for safety and probiotic potential. Its virulence and antibiotic resistant phenotypes (cytolysin and gelatinase production, antibiotic susceptibility) and genes (cylA, gelE, ace, agg, esp, and vanA) were surveyed. Results indicated that the tested virulence determinants were nontoxic. In addition, E. faecium YF5 was sensitive to 3 antibiotics such as amoxicillin, vancomycin, and chloramphenicol. Furthermore, results of in vivo animal acute oral toxicity of E. faecium YF5 studies were similar to the control group that indicated no abnormalities. In addition, E. faecium YF5 stably survived in low pH, bile salts, gastric, and intestinal fluids in vitro. Moreover, E. faecium YF5 was found to adhere to human colon cancer cell line HT‐29 at 3.39 (±0.67) × 10⁵ CFU/mL. When cocultured with pathogenic organisms (Enterobacter sakazakii CMCC45402, Escherichia coli CMCC44102, enterohemorrhage Escherichia coli O157: H7 CMCC44828, Salmonella Typhimurium CMCC50071, Shigella flexneri 301, and Shigella sonnei ATCC 29930) and 2 gram‐positive strains (Listeria monocytogenes CMCC54001 and Staphylococcus aureus CMCC 26003), it inhibited these foodborne pathogens with exception of S. aureus. Therefore, E. faecium YF5 can be regarded as a safe strain and it may be used as a probiotic preparation or for microecologics.     

Inhibition kinetics of polyphenol oxidase by glutamic acid

The inhibition of polyphenol oxidase (PPO) by glutamic acid was investigated. Application of different concentrations of glutamic acid to mushroom solution and Ocimum basilicum L. extract showed that glutamic acid appeared to be an effective browning inhibitor. Glutamic acid showed uncompetitive inhibition for mushroom and Ocimum basilicum L. polyphenol oxidases using 4-methylcatechol as a substrate, for mushroom PPO using catechol as a substrate and for Ocimum basilicum L. polyphenol oxidase using pyrogallol as a substrate; mixed-type inhibition for mushroom polyphenol oxidase using pyrogallol as a substrate; and non-competitive inhibition for Ocimum basilicum L. polyphenol oxidase using catechol as a substrate. Furthermore, sodium azide was used as an inhibitor for comparison with the inhibition potency of glutamic acid. It was found that glutamic acid was a more power inhibitor than sodium azide. The type of inhibition observed depended on the substrate, inhibitor and enzyme source used.     

Comparative essential oil composition of flowers,leavesand stems of basil (Ocimum basilicum L.) used as herb

The chemical composition of flower, leaves and stems from basil (Ocimum basilicum L.) have been examined by GC and GC–MS. The identified components constituting 99.03%, 95.04% and 97.66% of the flower, leaves and stem oils, respectively. The main constituents of the essential oil of flower, leaves and stem oils, respectively, were estragole (58.26%, 52.60% and 15.91%) and limonene (19.41%, 13.64% and 2.40%) and p-cymene (0.38%, 2.32% and 2.40%). Dill apiole (50.07%) was identified as the highest main constituent for stem. Estragole (15.91%), apiole (9.48) and exo-fenchyle acetate (6.14%) followed in order to decreasing them. Minor qualitative and major quantitative variations for some compounds of essential oils were determined with respect to different parts of O. basilicum. It was reported that the chemical composition of different parts oils of basil are very variable. It is known that specific estragole chemotypes are also known.     

Chemical composition of Ocimum basilicum L. essential oil and its efficacy as a preservative against fungal and aflatoxin contamination of dry fruits

The study presents fungal and aflatoxin contamination of some dry fruits and Ocimum basilicum essential oil (EO) as a plant‐based preservative. During mycoflora analysis, 2045 fungal isolates were recorded from dry fruits and 40% isolates of Aspergillus flavus were toxigenic in nature. The EO of O. basilicum exhibited strong fungitoxicity against toxigenic strain of A. flavus. Its minimum inhibitory concentration (MIC) was recorded at 1.0 μL ml^?1, and it completely inhibited aflatoxin B₁ production at 0.5 μL ml^?1. The oil exhibited broad fungitoxic spectrum and considerably reduced A. flavus isolates from dry fruits when used as fumigant in closed storage containers at 1.0 μL ml^?1. The chemical profile of the EO was standardised through GC–MS analysis. Based on antifungal potency, antiaflatoxigenicity and efficacy as fumigant during storage conditions, O. basilicum EO may be recommended as a botanical preservative for enhancing the shelf life of dry fruits and edible products during storage.     

Formation and dispersal of biofilms in dairy substrates

The formation and dispersion of biofilms in the dairy industry is a problem because it increases cross‐contamination, affecting the shelf life of the products and their safety. The objective of this study was to evaluate the influence of different dairy substrates (cows’ milk and whey protein) on the formation and dispersion of Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus and Bacillus cereus in two biofilm systems (mono‐species and multi‐species) on stainless steel at 25 °C. The dominant behaviour of E. faecalis occurred in most of the tests on mono‐species and multi‐species biofilms. A greater dispersion of biofilm cells was observed in skimmed milk.     

Development of PLA films containing oregano essential oil (Origanum vulgare L. virens) intended for use in food packaging

Consumers’ concerns about the environment and health have led to the development of new food packaging materials avoiding petroleum-based matrices and synthetic additives. The present study has developed polylactic acid (PLA) films containing different concentrations of essential oil from Origanum vulgare L. virens (OEO). The effectiveness of this new active packaging was checked for use in ready-to-eat salads. A plasticising effect was observed when OEO was incorporated in PLA films. The rest of the mechanical and physical properties of developed films did not show much change when OEO was included in the film. An antioxidant effect was recorded only for films containing the highest percentages of the active agent (5% and 10%). In addition, films exhibited in vitro antibacterial activity against Staphylococcus aureus, Yersinia enterocolitica, Listeria monocytogenes, Enterococcus faecalis and Staphylococcus carnosus. Moreover, in ready-to-eat salads, antimicrobial activity was only observed against yeast and moulds, where 5% and 10% of OEO was the most effective.     

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.     

Essential oil composition and antibacterial activity of the grapefruit (Citrus Paradisi. L) peel essential oils obtained by solvent‐free microwave extraction: comparison with hydrodistillation

The chemical composition of the essential oil (EO) obtained by solvent‐free microwave extraction (SFME) and hydrodistillation (HD) from the peel of grapefruit (Citrus Paradisi. L) was analysed by gas chromatography/mass spectrometry (GC/MS). Totally, twenty‐five components were identified in the EO. Limonene was observed as dominant (91.5–88.6%) for two extraction methods, SFME and HD, respectively. β‐Pinene (0.8–1.2%), linalool (1.1–0.7%), α‐terpinene (0.7–1.0%) and the other minor components were also detected. Disc diffusion method was applied to determine the antibacterial properties of the EO. The results showed that the EO of grapefruit peel had a wide spectrum of antimicrobial activities against Staphylococcus aureus, Enterococcus faecalis, Staphylococcus epidermidis, Escherichia coli, Salmonella typhimurium, Serratia marcescens and Proteus vulgaris, with their inhibition zones ranging from 11 to 53 mm.     

Dissemination of Enterococcus faecalis and Enterococcus faecium in a Ricotta Processing Plant and Evaluation of Pathogenic and Antibiotic Resistance Profiles

In this work, the sources of contamination by Enterococcus spp. in a ricotta processing line were evaluated. The isolated strains were tested for virulence genes (gelE, cylA,B, M, esp, agg, ace, efaA, vanB), expression of virulence factors (hemolysin and gelatinase), and the resistance to 10 different antibiotics. Enterococcus faecium and Enterococcus faecalis were subjected to discriminatory identification by intergenic spacer region (ITS)‐polymerase chain reaction and sequencing of the ITS region. The results showed that Enterococcus spp. was detected in the raw materials, environment samples and the final product. None of the 107 Enterococcus isolates were completely free from all virulence genes considered. A fraction of 21.5% of the isolates containing all of the genes of the cylA, B, M operon also expressed β‐hemolysis. Most of the isolates showed the gelE gene, but only 9.3% were able to hydrolyze gelatin. In addition, 23.5% of the observed Enterococcus isolates had the vanB gene but were susceptible to vancomycin in vitro. The dissemination of antibiotic‐resistant enterococci was revealed in this study: 19.3% of the E. faecium samples and 78.0% of the E. faecalis samples were resistant to at least one of the antibiotics tested. Sequencing of region discriminated 5 and 7 distinct groups among E. faecalis and E. faecium, respectively. Although some similarity was observed among some of the isolates, all E. faecalis and E. faecium isolates had genetic differences both in the ITS region and in the virulence profile, which makes them different from each other.     

Evaluation of antimicrobial and proteolytic activity of enterococci isolated from fermented products

A collection of 26 enterococci isolated from dairy and meat products were tested for antimicrobial and proteolytic activity. Enterococcus faecium and E. faecalis were the most frequent species among tested enterococci, and 11 isolates produced antimicrobial compounds. Results revealed that 10 out of 11 enterococci synthesized enterocins showing antimicrobial activity against food-born pathogen such as Listeria monocytogenes and Staphylococcus aureus. The broadest spectrum of antimicrobial activity was detected in E. faecalis BGPT1-10P and BGPT1-78. E. faecalis BG221 showed antimicrobial activity that was not related to production of enterocin, H₂O₂ or organic acid. Twenty-five enterococci showed strong or moderate proteolytic activity towards β-casein. Two isolates, BGPT1-10P and BGPT1-78, showed the most intense hydrolysis of α_s1-, β-, κ-casein fractions, total casein as well as gelatin. Extracellular BGPT1-10P and BGPT1-78 proteinases have a molecular mass of about 29 kDa. Bacteriocin production and proteinase activity of natural isolates of enterococci may be of technological interest in dairy and meat-fermented products.