Essential oils composition,antioxidant and antimicrobial activity of the leaves and flowers of Chaerophyllum macropodum Boiss

This study reports the essential oils chemical composition and in vitro antioxidant and antimicrobial potentials of the leaves and flowers of Chaerophyllum macropodum. GC and GC/MS analysis of the plant essential oils led to the identification of 49 components making 98.3–99.4% of its oils. The main constituents of the essential oils were trans-β-farnesene, trans-β-ocimene, β-pinene, limonene, spathulenol and myrcene constituting 49.6–73.1% of the oils. The extracts from the leaves and flowers showed moderate antioxidant activities in 1,1-diphenyl-2-picrylhydrazyl (DPPH) test (IC₅₀ values = 196.8 and 167.1 μg/ml respectively) and β-carotene/linoleic acid assay (inhibitions percentages = 69.9% and 62.7% respectively), but the essential oils were almost inactive in these tests. On the other hand, only the essential oils of the plant showed considerable antimicrobial activity against most of the tested microorganisms.     

Curcuma longa L. essential oil composition,antioxidant effect,and effect on Fusarium verticillioides and fumonisin production

Essential oils may be an alternative to the use of synthetic fungicides for the control of fungi involved in agricultural product deterioration. The aim of the present study was to evaluate the composition and antioxidant effect of turmeric essential oil and its antifungal and antimycotoxigenic action on Fusarium verticillioides (Sacc.) Nirenberg. The essential oil major components were α-turmerone (42.6%), β-turmerone (16.0%) and ar-turmerone (12.9%). The half-maximal inhibitory concentration (IC₅₀) for the radical scavenging capacities of 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were 0.54 and 10.03 mg/ml, respectively, indicating good antioxidant activity. The application of 17.9 and 294.9 μg/ml of turmeric essential oil decreased the development of F. verticillioides by 56.0 and 79.3%, respectively, when compared with the fungal control. The scanning electron microscopy demonstrated that the oil decreased the thickness and the length of the microconidia. Ergosterol production significantly decreased (p < 0.05) in groups treated with the essential oil relative to the control, indicating an effect of the oil on fungal biomass. The production of B₁ and B₂ fumonisins was significantly inhibited (p < 0.05) in groups treated with the essential oil. The results suggest that turmeric essential oil has antioxidant, antifungal and antimycotoxigenic activities.     

Antimicrobial and antioxidant activities of Chinese sumac (Rhus typhina L.) fruit extract

Sumac plant (Rhus typhina) is mainly used for forestation and gardening, whereas its fruit can be used to prepare a kind of beverage called “sumac-ade” and to treat gastrointestinal disorders. This study aimed at assaying the antimicrobial and antioxidant activities of R. typhina growing in China. The antimicrobial activity of R. typhina fruit extract was tested against twelve strains including Gram-positive and Gram-negative bacteria as well as yeasts. The extract showed a strong antimicrobial activity with a concentration-dependence and a broad antimicrobial spectrum for all tested bacteria species. Bacillus cereus and Helicobacter pylori were found to be the most sensitive Gram-positive and Gram-negative bacteria respectively, with a minimum inhibitory concentration (MIC) of 0.10%. However, yeasts exhibited much lower sensitivity, with MICs of 0.60–0.75%. Furthermore, the antioxidant activity of the extract was investigated, including scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals (0.016 mg/ml as IC₅₀ value) and reducing power (IC₅₀ value of 0.041 mg/ml). To our knowledge, this is the first report on the biological activities of R. typhina fruit extract and our findings suggest the possibility of using the fruit of R. typhina as a novel source of natural antimicrobial and antioxidant agents for the food and pharmaceutical industries.     

Chemical composition and in vitro antimicrobial,antifungal and antioxidant properties of essential oils obtained from some herbs widely used in Portugal

The aim of this study was determine (i) the chemical composition (ii) the antimicrobial activity (antibacterial and antifungal) and (iii) the antioxidant activity by means of four different antioxidant tests (DDPH, FIC, FRAP and TBARS) of the EOs of three aromatic herbs, Coriander (Coriandrum sativum), celery (Apium graveolens) and bush-basil (Ocimum minimum) widely used in Portugal.There is a great variability of the compounds presented in the three tested essential oils. Bush-basil EO had the highest total phenolic content (794.9 mg GAE/L) while coriander EO had the lower total phenolic content (52.3 mg GAE/L). Since bush-basil had the highest TPC it was expected to present a very high antioxidant profile, which was verified in 3 of the 4 assays (DPPH inhibition of 95.9%; FRAP values of 2.7 mmol Trolox/L; TBARS inhibition of 87.2%); coriander, despite the low TPC showed the highest inhibition in the FIC assay (94.1%).The bush-basil EO showed the highest antimicrobial activity, with MIC ranging between 0.6 and 5 μL/mL against bacteria and 0.04–2.5 μL/mL against yeasts. Both celery and coriander EO had a very similar antimicrobial activity against all the tested strains. The antifungal activity was higher in the bush-basil EO against Mucor racemosus and Penicillium chrysogenum since it was the only EO that showed growth inhibition on all the tested concentrations. Alternaria alternata showed great resistance against all the tested essential oils.     

In vitro evaluation of antibacterial and antioxidant activities of the essential oil and methanol extract of endemic Zataria multiflora Boiss

The present study was conducted to evaluate in vitro antibacterial and antioxidant properties of essential oil and methanol extracts from a unique and endemic plant, Zataria multiflora Boiss. The antibacterial test results showed that the essential oil of the plant strongly inhibited the growth of all of the microorganisms studied especially the Gram-negative strains. The polar fraction of methanol extract has been effective against Gram-positive strains, while the non-polar fraction has shown activity similar to essential oil. The antioxidant potential of the samples was evaluated using two separate methods, inhibition of free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ammonium thiocyanate systems. Sub fractions of the methanol extract were able to reduce the stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) with an IC₅₀ of 11.7 ± 1.58 and 16.2 ± 1.61 μg/ml, respectively for non-polar and polar ones, which the activity of the latter almost is equal to synthetic antioxidant BHA (18.2 ± 1.94 μg/ml). Inhibition values of linoleic oxidation were calculated to be 82.4% and 80.3% for the polar and non-polar fractions, respectively. The essential oil to be showed more inhibition (89.7 ± 2.5), similar to the synthetic antioxidants BHA (97.8 ± 2.94) and ascorbic acid (93.2 ± 2.1). The chemical composition of hydrodistilled essential oils of Z. multiflora was analyzed by GC/MS. A total of 25 compounds representing 99.78% of the oil were identified: thymol (37.59%), carvacrol (33.65%); para-cymene (7.72%), γ-terpinene (3.88%) and β-caryophyllene (2.06%) were the main components comprising 84.9% of the oil. Results here show that the essential oil and methanol extract of Z. multiflora possess antioxidant and antibacterial activity, and therefore it could be used as a natural preservative ingredient in food and/or pharmaceutical industries.     

In vitro assessment of synthetic phenolic antioxidants for inhibition of foodborne Staphylococcus aureus,Bacillus cereus and Pseudomonas fluorescens

Minimal inhibitory concentrations (MICs) of seven synthetic phenolic compounds, five commonly used as antioxidants (TBHQ, BHA, BHT, propyl gallate and octyl gallate) and two as antimicrobials (propyl 4-hydroxybenzoate and n-heptyl 4-hydroxybenzoate) were assessed against several strains of two Gram-positive (Staphylococcus aureus and Bacillus cereus) and one Gram-negative (Pseudomonas fluorescens) bacteria, by using a standardized microdilution assay (ISO 20776-1, 2006). Octyl gallate was the most effective compound against the three genera/species of bacteria considered simultaneously (with the exception of four strains of B. cereus, which were resistant for this compound) with MIC values (≤100 μg/ml) lower than the concentrations usually used as antioxidants. TBHQ and n-heptyl 4-hydroxybenzoate were also effective in the control of S. aureus at very low concentrations (MIC of 3.1 μg/ml and 12.5 μg/ml, respectively). Propyl 4-hydroxybenzoate was the most inhibitory phenolic compound against all strains of B. cereus and both tested parabens (propyl- and heptyl-) were not effective for P. fluorescens (MIC > 1600 μg/ml). B. cereus was the bacterial genera that showed more intra-species variation, distinguishing two clearly groups of sensitivity among the strains to octyl gallate and n-heptyl 4-hydroxybenzoate (“sensitive” with mean MICs of 42.8 and 4.2 μg/ml, respectively; and “resistant” with MICs >1600 and >800 μg/ml, respectively). According to all that, octyl gallate would be an interesting phenolic compound for the food industry, not only because of its recognized antioxidant properties but also because of their effectivity as antimicrobial against S. aureus, B. cereus and P. fluorescens.     

Chemical composition of commercial citrus fruit essential oils and evaluation of their antimicrobial activity acting alone or in combined processes

The present investigation reports on the chemical composition of three commercial citrus fruit essential oils (orange [Citrus sinensis], lemon [Citrus lemon] and Mandarin [Citrus reticulata]) from Spain, and examines their antimicrobial activity against spoiling and pathogenic microorganisms, as well as possible synergistic lethal effects in combination with mild heat. Gas chromatography-mass spectrometry analysis allowed for the identification of 65 compounds as main volatile constituents. Limonene was the major component in the three essential oils (59–85%). Preliminary results obtained by using the disc agar diffusion technique indicated mandarin EO as the best growth inhibitor and discarded orange and lemon essential oils. The evaluation of the bacteriostatic and bactericide effect confirmed the broadest spectrum of action to be that of the mandarin essential oil. However, low concentrations of the three essential oils (0.2 μl/ml) in combination with a mild heat treatment (54 °C/10 min) showed synergistic lethal effects, inactivating more than 5 log cycles of bacterial cells, and demonstrating their potential in the design of successful combined treatments for food preservation.     

Degradation of fumonisin B1 by cinnamon essential oil

Fumonisins are group of mycotoxins produced mainly by Fusarium verticillioides and Fusarium proliferatum. They frequently contaminate corn and corn based products, and cause several diseases in humans and animals. Fumonisin B₁ (FB₁) is the most prevalent fumonisin and is highly toxic to human and animal. The essential oils from plants offer a hope in the prevention and detoxification of these mycotoxins. The present study investigates the degradation effect of cinnamon, citral, Litsea cubeba, clove, eucalyptus, anise, spearmint and camphor oils on FB₁. The degradation level of FB₁ was determined by ELISA. Cinnamon oil proves to be effective essential oil in reducing FB₁, followed by citral, eugenol oil, eucalyptus oil, anise oil and camphor oil. The effects of incubation time, and temperature with respect to the concentration of cinnamon oil on their degradation effect on FB₁ by cinnamon oil were investigated. Results showed that at 120 h time with the 280 μg/ml concentration of cinnamon oil, under 30 °C is optimal for FB₁ reduction. Under optimal condition, FB₁ was reduced from 15.03 to 0.89 μg/ml (94.06%). Cinnamon oil could be a promising candidate in the detoxification and control of FB₁ in corn based products.     

Antioxidant activity of essential oil and aqueous extract of Pelargonium graveolens L’Her

Pelargonium graveolens L’Her is an aromatic, rose-scented herb which is indigenous to various parts of southern Africa, and nowadays cultivated worldwide. This work presents the first phytochemical investigation of P. graveolens cultivated in Bosnia. The volatile profile of odorous parts of the plant was analysed by GC/MS. More than eighty compounds were identified in essential oils obtained from the leaves and stems, representing 92.3% and 96.3% in total, respectively. The major compounds in essential oils were oxygenated monoterpenes (64.3-74.2%), with geraniol (27.5-50.2%) and citronellol (14.2-19.0%) as the main representatives. The content of phenolic compounds in corresponding hydrosols as 34.88 ± 2.00 mg GAE/g in leaves and 102.44 ± 1.63 mg GAE/g in stems including flavonoid compounds of 32.35 ± 0.81 mg GAE/g to 101.87 ± 1.03 mg GAE/g. The radical-scavenging activity was measured by the DPPH method and IC₅₀ values ranged from 0.19 ± 0.05 mg/mL (stems) to 0.39 ± 0.04 mg/mL (leaves) for hydrosols, and from 63.70 ± 1.56 mg/mL (leaves) to 64.88 ± 1.12 mg/mL (stems) for essential oils.     

Effects of extraction methods of shiitake by-products on their antioxidant and antimicrobial activities in fermented sausages during storage

Two different solvents (water and ethanol/water at 50/50 ratio) were used for extraction of phenolic contents in shiitake by-products, and the antioxidant and antimicrobial efficacies of fermented sausages incorporated with these two extracts were investigated during storage. Also, shelf-life stability of fermented sausages fortified with these two extracts was compared with those fortified with BHT (0.02%) or nitrite (0.01%)/nitrate (0.005%). Significantly higher antioxidant activities (e.g., ferrous ion chelating and DPPH scavenging) and inhibitory capacity against lipid oxidation (e.g., TBARS reduction) were observed in the fermented sausages made with ethanolic extract (shiitake by-products extracted with ethanol/water) compared with those added with aqueous extract (extracted with water) or BHT, nitrite/nitrate and control throughout storage. The ethanolic extract treated-samples also showed a significantly slower increasing rate of total aerobic count (6.54–6.95 log₁₀ cfu/g) than the ones treated with aqueous extract (6.74–7.16 log₁₀ cfu/g) during storage. Otherwise, extract obtained from the ethanolic extract treated-samples had stronger antimicrobial activities against pathogens than the one obtained from aqueous extract treated-samples (e.g., minimum inhibitory concentrations, MIC = 2.05 & 3.64, 3.12 & 5.20 and 7.29 & 10.41 mg/mL for S. aureus, L. monocytogenes and E. coli O157, respectively). Especially, the antimicrobial activity against S. typhimurium (MIC = 37.50 mg/mL) was observed only in the extract of ethanolic extract treated-samples. Our study demonstrates that the extraction with ethanol/water solvent is a more effective method to obtain bioactive compounds enriched-extract which better improved the shelf-life stability of fermented sausages during storage without defects in quality in comparison to the extraction method with water.     

Determination of antimicrobial effect of mint and basil essential oils on survival of E. coli O157:H7 and S. typhimurium in fresh-cut lettuce and purslane

In this research, mint and basil essential oils at concentrations of 0.01 ml/L, 0.032 ml/L or 0.08 ml/L were used for disinfection treatments of fresh-cut lettuce and purslane samples inoculated with Salmonella typhimurium or Escherichia coli 0157:H7. Disinfection treatment time was applied as 10 min (short) or 15 min (long). Disinfected samples were packaged aerobically and stored at refrigerator +4 °C for 7 days. It was observed that mint and basil essential oils showed antimicrobial effect on the survival of E. coli O157:H7 and S. typhimurium inoculated into lettuce and purslane samples during refrigerated storage. Mint essential oil showed higher antimicrobial effect on pathogens when compared to basil essential oil. Mint and basil essential oils at concentration of 0.08 ml/L were the most effective antimicrobial treatment against pathogens in two different vegetable. S. typhimurium was more resistant against basil oil in lettuce samples when compared to its resistance against basil oil in purslane samples, whereas E. coli O157:H7 was more resistant to mint oil in purslane samples when compared to its resistance against mint oil in lettuce samples.     

Novel biocompatible silver nanoparticles for controlling the growth of lactic acid bacteria and acetic acid bacteria in wines

This paper reports the inhibitory potential of two new silver-based biocompatible nanoparticles (Ag NPs), specifically synthesized for this study, against Gram-negative and Gram-positive bacteria, such as Escherichia coli and Staphylococcus aureus, and different wine lactic acid bacteria (LAB) (Oenococcus oeni, Lactobacillus casei, Lactobacillus plantarum and Pediococcus pentosaceus) and acetic acid bacteria (AAB) (Acetobacter aceti and Gluconobacter oxydans). Both solid nanoparticles, dried PEG-Ag NPs 1 (20.01% silver content) and glutathione-stabilized GSH-Ag NPs 2 solution (0.197 mg/mL silver content), showed homogeneous size distribution and physicochemical properties compatible with their use as antimicrobial agents. The nanoparticles showed different antimicrobial spectra, with PEG-Ag NPs 1 being more effective (lower IC₅₀ values) against Gram-negative strains, whereas GSH-Ag NPs 2 showed similar efficiency against Gram-negative and Gram-positive strains, with the exception of O. oeni strains, which turned out to be notably susceptible to them (IC₅₀ ∼1 μg/mL, Ag concentration). For the LAB and AAB tested, these nanoparticles showed IC₅₀ values (particle concentration) of the same order as those for the widely used additive potassium metabisulphite, at least as calculated for PEG-Ag NPs 1. These results confirm the potential of Ag NPs in controlling microbial processes in winemaking, and open new investigations on the design and use of antimicrobial-specific silver-based nanoparticles in oenology.     

Synergistic action between fractions of essential oils from Cymbopogon citratus, Ocimum gratissimum and Thymus vulgaris against Penicillium expansum

Penicillium expansum is a mould that causes the rotting of several fruits and vegetables, especially apples onto which it also synthesizes some dangerous mycotoxins. The degree of synergism between fractions of essential from Cymbopogon citratus, Ocimum gratissimum and Thymus vulgaris was evaluated against two mycotoxin producing strains of P. expansum. The antifungal activity determined by dilution method and expressed as a Number of Decimal Reduction of the colony forming units per ml (NDR cfu) showed that the essential oils extracted from O. gratissimum was significantly (P < 0.05) more active against P. expansum than those extracted from C. citratus and T. vulgaris. Fractions enriched with oxygenated terpenes were significantly (P < 0.05) more active than their respective essential oils, whereas most of the fractions enriched with terpene hydrocarbons, were significantly (P < 0.05) less active. The fungicidal activity of mixtures of fractions from the same essential oils or from two different essential oils showed that there exist synergistic, additive and antagonistic effects between fractions of the three essential oils tested against both fungal strains. The synergistic effects observed could be exploited in order to maximize the antimicrobial activity of essential oils and to minimize the concentrations of essential oil required to produce a given antimicrobial effect without any alteration of the food test.     

Biological activities of Boswellia sacra extracts on the growth and aflatoxins secretion of two aflatoxigenic species of Aspergillus species

Aflatoxins are the most serious carcinogenic, hepatotoxic, teratogenic and mutagenic secondary metabolites which adversely affect human and animal health. This study was designed to evaluate the in vitro inhibitory effect of different concentrations of Boswellia sacra resin (2.5, 5, 7.5 and 10 g/100 ml), leaf extract (5, 7.5, 10, 12.5 and 15 ml/100 ml), and essential oil (1, 2, 3, and 4 ml/100 ml) on the growth and aflatoxins production by two species of Aspergilli, namely Aspergillus flavus (SQU21) and Aspergillus parasiticus (CBS921.7). Resin of B. sacra caused 57.9–92.1% inhibition of aflatoxin secretion by A. flavus and 43.6–95.7% for A. parasiticus. However, the mycelial dry weights were significantly increased by 20.9–52.7% for A. flavus, and 8.9–68.5% for A. parasiticus. The leaf extract of B. sacra apparently enhanced aflatoxins production by 20–50%, and mycelial dry weight by 25.5–29.1% for A. flavus and A. parasiticus. The essential oil of B. sacra at different concentrations similarly inhibited the fungal growth and aflatoxins production by 45.8–83.7% for A. flavus and 41.3–83.5% for A. parasiticus which indicates the antifungal activity of this oil. None of the B. sacra extracts detoxified pure aqueous aflatoxin B₁. We have concluded that B. sacra resin and essential oil possess biological activity against biochemical synthesis and metabolic pathway of aflatoxin production of the two Aspergillus species. Therefore, the resin and essential oil of B. sacra can be recommended as safe plant based bioreservatives to enhance shelf life of food and feed products with reference to adverse effect of physical and synthetic chemical preservatives and their antimicrobial and aflatoxins inhibition activity.     

Immunochemical detection of apramycin as a contaminant in tissues of edible animals

Veterinary aminoglycoside antibiotic apramycin (AP) was used as hapten in different conjugation procedures for preparation of immunogen and coating antigens. Among the coating antigens one was chosen to ensure strictly selective determination of AP with no cross-reactivity of related analogs. The limit of competitive ELISA detection was 0.015 ng/ml for swine kidney and bovine muscle extracts. The dynamic range IC₂₀–IC₈₀ was calculated as 0.03–1.8 ng/ml (0.15–9 μg/kg). High sensitivity of assay allowed eliminating matrix effect of samples by simple dilution of tissue extracts. The experiments with tissue samples fortified with AP at half, single and double MRL values showed the recovery rate of 85–105%. Along with the other immunotests and kits for determination of aminoglycosides this for the first time developed assay of apramycin is suitable for screening of antibiotic residues in foodstuff.     

Evaluation of the antimicrobial and antioxidant activities of essential oils, extracts and their main components from oregano from Madeira Island, Portugal

Origanum vulgare subsp. virens growing wild in Madeira Island, Portugal was studied within the ongoing investigations on polymorphic Lamiaceae species. The antimicrobial activity of the essential oils and n-hexane extracts of origanum was determined against 10 strains of bacteria and yeasts, found as human pathogenic or food spoilage microorganisms. The essential oils, n-hexane extracts and isolated compounds showed moderately activity, compared to standard antibiotics, inhibiting all tested bacteria except Pseudomonas aeruginosa. The most sensitive microorganism was Mycobacterium smegmatis, reaching MIC = 25 μg mL⁻¹. The results obtained suggest a potential application of these oils in preventing the human pathogenic and food spoilage due to microorganism’s growth. The essential oils and n-hexane extracts have greater RSC than polar extracts, probably due to the high contents in thymol, which demonstrated the highest activity in the DPPH assay. All studied origanum samples showed a large content in non-esterified 1-hexacosanol, C₂₆H₅₄O, accumulated mainly in bracts and flowers.     

Preservative effectiveness of essential oils in vapor phase combined with modified atmosphere packaging against spoilage bacteria on fresh cabbage

This study was conducted to investigate the antibacterial effects of various essential oils (EOs) against pathogens using the disc volatilization method. Also, combined effects of EOs in vapor phase and MAP were evaluated for reducing levels of total mesophilic microorganisms on fresh cabbage. The vapor phase activities of EOs (thyme-1, oregano-1, lemongrass-1, and lemongrass-2 oils) observed strong inhibitory effects. The MAP results showed that 100% CO₂ gas packaging reduced significantly levels of total mesophilic microorganisms on cabbage and radish sprouts, and their reduction level was 1.55 and 2.26 log₁₀ CFU/g compared to control after 21 days of storage (p ≤ 0.05). Based on previous results, combined effects of EOs in vapor phase and MAP (100% CO₂) showed that lemongrass-2 oil with 20 discs showed complete inactivation by <1.0 log₁₀ CFU/g after 14 days of storage. These results could provide useful information for developing alternative preservation method to improve the freshness and shelf-life of fresh produce using natural antimicrobials.     

Chemical composition and antiaflatoxigenic activity of Carum carvi L., Thymus vulgaris and Citrus aurantifolia essential oils

In order to find out plants useful to controlling aflatoxins (AFs) production, the essential oils (EOs) from 12 medicinal plants prepared by hydrodistillation were studied with special reference to the inhibition of Aspergillus parasiticus growth and AFs production. The toxigenic fungus was cultured in presence of various oils in 6-well microplates using a microbioassay technique. The mycelial mass was estimated as an index of fungal growth, while the aflatoxins B₁ (AFB₁) and G₁ (AFG₁) were measured by high performance liquid chromatography (HPLC). Among plants tested, Thymus vulgari and Citrus aurantifolia were found to inhibit both A. parasiticus and AF production. The EOs from Mentha spicata L., Foeniculum miller, Azadirachta indica A. Juss, Conium maculatum and Artemisia dracunculus were only inhibited fungal growth, while Carum carvi L. effectively inhibited AF production without any obvious effect on fungal growth. The other plants including Ferula gummosa, Citrus sinensis, Mentha longifolia and Eucalyptus camaldulensis had no effect on A. parasiticus growth and AF production at all concentrations used. The IC₅₀ values of T. vulgaris, C. aurantifolia and C. carvi for AF inhibition were reported as 93.5, 285.6, and 621.9 μg/ml for AFB₁, while they were calculated as 11.7, 50.1, and 56.0 μg/ml for AFG₁. These results indicate that the EOs of some medicinal plants may be considered as potential candidates to protect foods and feeds from toxigenic fungus growth and subsequent AF contamination.     

Organic thyme oil emulsion as an alternative washing solution to enhance the microbial safety of organic cantaloupes

Emulsions of organic essential oils may be used as postharvest alternative washing solutions in fresh produce production. In the present study, organic thyme oil was emulsified with whey protein concentrate, gum arabic, lecithin, or their equal mass mixtures without using specialized equipment. The stability of these emulsions was monitored by measuring hydrodynamic diameter during ambient storage up to 7 days. The antimicrobial activity of these emulsions against Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). All emulsions had lower MIC and MBC than free thyme oil pre-dissolved in ethanol. Thyme oil emulsified with gum arabic had the smallest and most stable hydrodynamic diameter (156–239 nm) and was chosen as the washing solution to evaluate its efficacy in reducing pathogens on organic cantaloupes. Cantaloupes inoculated with pathogens were immersed in 0.1%, 0.2%, or 0.5% emulsified or free thyme oil for 2 min. The counts of the three cultures inoculated on cantaloupes were reduced by either 0.2% or 0.5% thyme oil and the emulsions were more effective than free thyme oil (P < 0.05). Organic load (2% or 5%) had no effect on their antimicrobial efficacy (P > 0.05). During ambient storage (21 °C) up to 10 days, the counts for all three bacteria gradually declined for all treatments and the emulsion treatment had consistently lower populations than unwashed and water-washed treatments. Therefore, emulsions of organic essential oils have potential applications as postharvest washing solutions to improve the microbiological safety of organic fresh produce.     

Effect of detergents as antibacterial agents on biofilm of antibiotics-resistant Vibrio parahaemolyticus isolates

Vibrio parahaemolyticus (V. parahaemolyticus) is a halophilic, Gram-negative human pathogen known as a leading cause of seafood-derived food poisoning. Due to high contamination rate of seafood in Asian countries, V. parahaemolyticus is considered as a food safety concern. V. parahaemolyticus is able to produce biofilm which is more resistant toward disinfectants and antibodies than its planktonic form. Thirty six V. parahaemolyticus isolates from seafood were tested for their susceptibility using 18 different antibiotics. Two V. parahaemolyticus isolates were resistant to bacitracin, chloramphenicol, rifampin, ampicillin, vancomycin, nalidixic acid, penicillin and spectinomycin. Fourteen V. parahaemolyticus isolates were found to be resistant to bacitracin, tetracycline, rifampin, ampicillin, vancomycin, penicillin and spectinomycin. The remaining two isolates were resistant to more than 2 antibiotics. Majority of the V. parahaemolyticus isolates (97.2%) showed MAR index > 0.2, indicating that these isolates were originated from high risk sources. To investigate effect of three common detergents on antibacterial-resistant V. parahaemolyticus, 16 V. parahaemolyticus isolates resistant to more than 7 antibiotics were selected. V. parahaemolyticus (ATCC 17802) was used as reference strain. Detergents were tested for their minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) and time–kill curves were constructed to assess the concentration between MIC and bactericidal activity of detergents. Detergents D1 (Linear alkyl benzene based) was found to be the most effective with MIC and MBC ranged between 97.656 and 1562.5 μg/ml and 781.25–3125 μg/ml, respectively. The time–kill curves demonstrated that the bactericidal endpoint for resistant V. parahaemolyticus isolates reached after 30 min incubation with D1 at concentration 8 × MIC. The isolate VP003 was killed at 8 × MIC within 0.5 h and the reduction in CFU/ml was 3 log units (99.9%). V. parahaemolyticus biofilms were formed in 96 wells microtiter plates at 37 °C and 24 h-old biofilm were used to test antibacterial activity of detergents. Results showed that biofilm-producing ability of antibacterial-resistant V. parahaemolyticus isolates were inhibited at 1562.5–6250 μg/ml of D1 and eradicated at 3125 – ≥50,000 μg/ml of D1. Detergents showed potential antimicrobial activity against V. parahaemolyticus