Exploring the Diverse Biological Properties of Cannabidiol: A Focus on Plant Growth Stimulation.

The aim of the current study was to compare some biological activities of edible oils enriched with 10 % of cannabidiol (CBD samples) from the Slovak market. In addition, hemp, coconut, argan, and pumpkin pure oils were also examined. The study evaluated the fatty acids content, as well as antibacterial, antifungal, antioxidant, cytotoxic, and phytotoxic activities. The CBD samples presented antimicrobial activity against the tested bacterial strains at higher concentrations (10000 and 5000 mg/L) and antifungal activity against Alternaria alternata, Penicillium italicum and Aspergillus flavus. DPPH⋅ and FRAP assays showed greater activity in CBD-supplemented samples compared to pure oils and vitamin E. In cell lines (IPEC-J2 and Caco-2), a reduced cell proliferation and viability were observed after 24 hours of incubation with CBD samples. The oils showed pro-germinative effects. The tested activities were linked to the presence of CBD in the oils.

Novel Antifungal Dimers from the Roots of Taiwania cryptomerioides.

Five new dimer compounds, namely Taiwaniacryptodimers A-E (1-5), were isolated from the methanol extract of the roots of Taiwania cryptomerioides. Their structures were established by mean of spectroscopic analysis and comparison of NMR data with those of known analogues. Their antifungal activities were also evaluated. Our results indicated that metabolites 1, 2, 4, and 5 displayed moderate antifungal activities against Aspergillus niger, Penicillium italicum, Candida albicans, and Saccharomyces cerevisiae.

Epilobium angustifolium L. Essential Oil-Biological Activity and Enhancement of the Skin Penetration of Drugs-In Vitro Study.

Epilobium angustifolium L. is a popular medicinal plant found in many regions of the world. This plant contains small amounts of essential oil whose composition and properties have not been extensively investigated. There are few reports in the literature on the antioxidant and antifungal properties of this essential oil and the possibility of applying it as a potential promoter of the skin penetration of drugs. The essential oil was obtained by distillation using a Clavenger type apparatus. The chemical composition was analyzed by the GC-MS method. The major active compounds of E. angustifolium L. essential oil (EOEa) were terpenes, including α-caryophyllene oxide, eucalyptol, ß-linalool, camphor, (S)-carvone, and ß-caryophyllene. The analyzed essential oil was also characterized by antioxidant activity amounting to 78% RSA (Radical Scavenging Activity). Antifungal activity against the strains Aspergillus niger, A. ochraceus, A. parasiticum, and Penicillium cyclopium was also determined. The largest inhibition zone was observed for strains from the Aspergillus group. The EOEa enhanced the percutaneous penetration of ibuprofen and lidocaine. After a 24 h test, the content of terpene in the skin and the acceptor fluid was examined. It has been shown that the main compounds contained in the essential oil do not penetrate through the skin, but accumulate in it. Additionally, FTIR-ATR analysis showed a disturbance of the stratum corneum (SC) lipids caused by the essential oil application. Due to its rich composition and high biological activity, EOEa may be a potential candidate to be applied, for example, in the pharmaceutical or cosmetic industries. Moreover, due to the reaction of the essential oil components with SC lipids, the EOEa could be an effective permeation enhancer of topically applied hydrophilic and lipophilic drugs.

Wood Extractives of Silver Fir and Their Antioxidant and Antifungal Properties.

The chemical composition of extractives in the sapwood (SW), heartwood (HW), knotwood (KW), and branchwood (BW of silver fir (Abies alba Mill.) was analyzed, and their antifungal and antioxidant properties were studied. In addition, the variability of extractives content in a centripetal direction, i.e., from the periphery of the stem towards the pith, was investigated. The extracts were analyzed chemically with gravimetry, spectrophotometry, and chromatography. The antifungal and antioxidative properties of the extracts were evaluated by the agar well diffusion method and the diphenyl picrylhydrazyl radical scavenging method. Average amounts of hydrophilic extractives were higher in KW (up to 210.4 mg/g) and BW (148.6 mg/g) than in HW (34.1 mg/g) and SW (14.8 mg/g). Extractives identified included lignans (isolariciresinol, lariciresinol, secoisolariciresinol, pinoresinol, matairesinol) phenolic acids (homovanillic acid, coumaric acid, ferulic acid), and flavonoids epicatechin, taxifolin, quercetin). Secoisolariciresinol was confirmed to be the predominant compound in the KW (29.8 mg/g) and BW (37.6 mg/g) extracts. The largest amount of phenolic compounds was extracted from parts of knots (281.7 mg/g) embedded in the sapwood and from parts of branches (258.9 mg/g) adjacent to the stem. HW contained more lignans in its older sections. Hydrophilic extracts from knots and branches inhibited the growth of wood-decaying fungi and molds. KW and BW extracts were better free radical scavengers than HW extracts. The results of the biological activity tests suggest that the protective function of phenolic extracts in silver fir wood can also be explained by their antioxidative properties. The results of this study describe BW as a potential source of phenolic extractives in silver fir.

A Comparative Study of Antimicrobial and Antioxidant Activities of Plant Essential Oils and Extracts as Candidate Ingredients for Edible Coatings to Control Decay in 'Wonderful' Pomegranate.

This study determined the antimicrobial and antioxidant activity of lemongrass (LO), thyme (TO), and oregano (OO) essential oils and ethanolic extracts of pomegranate peel (PPE) and grape pomace (GPE) as candidate ingredients for edible coatings. Antifungal effects against Botrytis cinerea and Penicillium spp. were tested using paper disc and well diffusion methods. Radical scavenging activity (RSA) was evaluated using 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid assays. Gas chromatography-mass spectrometry analysis identified limonene (16.59%), α-citral (27.45%), ß-citral (27.43%), thymol (33.31%), paracymene (43.26%), 1,8-cineole (17.53%), and trans-caryphellene (60.84%) as major compounds of the essential oils. From both paper disc and well diffusion methods, LO recorded the widest zone of inhibition against tested microbes (B. cinerea and Penicillium spp.). The minimum inhibitory concentrations of LO against B. cinerea and Penicillium spp., were 15 µL/mL and 30 µL/mL, respectively. The highest (69.95%) and lowest (1.64%) RSA at 1 mg/mL were recorded for PPE and OO. Application of sodium alginate and chitosan-based coatings formulated with LO (15 or 30 µL/mL) completely inhibited spore germination and reduced the decay severity of 'Wonderful' pomegranate. Lemongrass oil proved to be a potential antifungal agent for edible coatings developed to extend shelf life of 'Wonderful' pomegranate.

Control of postharvest diseases caused by Penicillium spp. with myrtle leaf phenolic extracts: in vitro and in vivo study on mandarin fruit during storage.

BACKGROUND: In the postharvest handling of horticultural commodities, plant extracts with fungicidal activity are a valid alternative to synthetic fungicides. The fungicidal activity of myrtle leaf extracts from eight cultivars was studied in vitro against Penicillium digitatum, Penicillium italicum, and Penicillium expansum and on artificially inoculated mandarins with green and blue molds during storage for 12 days at 20 °C and 90% RH. RESULTS: Hydroxybenzoic acids, hydrolysable tannins, and flavonols were identified by high-performance liquid chromatography (HPLC). Despite sharing the same phenolic profile, extracts of eight myrtle cultivars significantly differed in the concentration of phenolics. Hydrolysable tannins are the principal subclass representing nearly 44.9% of the total polyphenols, whereas myricitrin was the most abundant flavonol in all cultivars. Myrtle extracts strongly inhibited conidial germination of the pathogens tested, although the greatest efficacy was observed against P. digitatum. At a concentration of 20 g L-1 , all the extracts completely inhibited fungi growth; only 'Angela', 'Tonina' and 'Grazia' extracts were effective at lower concentrations (15 g L-1 ). On inoculated fruit, myrtle extracts significantly controlled rot development. As a preventive treatment, 'Ilaria' and 'Maria Rita' extracts significantly reduced the rate of fruit with green mold decay lesions. When applied as a curative treatment, all the exacts decreased the incidence of decay. Against P. italicum, all the extracts applied as preventive treatments controlled decay effectively, while as curative treatment some of the extracts were not effective. All the extracts reduced the size of the infected areas. CONCLUSION: The results propose myrtle extracts as a possible natural alternative to synthetic fungicides. © 2021 Society of Chemical Industry.

Toxicity and Antimicrobial Activities of Amaranthus caudatus L. (Amaranthaceae) Harvested From Formulated Soils at Different Growth Stages.

This study examined the toxicity and antimicrobial effects of ethanol and aqueous extracts from Amaranthus caudatus grown on soils formulated from parent particles of silt, sand and clay in a glasshouse. Four different soils namely; sandy clay loam, loam, clayey loam and silty clay loam from were formulated were used for cultivation with the unfractionated soil which was the control. Crude extracts obtained from the plant shoots harvested at different growth stages were tested on some certain gram-negative and gram-positive bacteria and some fungi via agar dilution assay. The toxicity of the water and ethanol extracts was also examined via Artemia salina assay and the level of lethality was measured against Clarkson's lethality scale. All aqueous samples, as well as ethanol extracts of flowering and pre-flowering harvests of control soil tested, were non-toxic (LC50 > 1 mg/mL). At post flowering, the ethanolic extracts were highly toxic mostly in clayey loam, control, sandy-clayey loam soils (LC50 < 0.5 mg/mL). Also, antifungal effects of the plant revealed that extracts inhibited the growth of Candida albicans significantly with mild effect on Candida glabrata, Penicillium chrysogenum and Penicillium aurantiogriseum suggesting that the plant is a promising pharmacological candidate in the treatment of candidiasis. For an optimal yield of non-toxic supplement for household consumption which may also serve as pharmacological precursors, clayey loam soil is recommended for cultivation and harvesting may occur at pre-flowering or flowering stage using ethanol and water as solvents of extraction.

Unravelling the Antifungal Effect of Red Thyme Oil (Thymus vulgaris L.) Compounds in Vapor Phase.

The aim of this work was to evaluate the antifungal activity in vapor phase of thymol, p-cymene, and γ-terpinene, the red thyme essential oil compounds (RTOCs). The Minimum Inhibitory Concentration (MIC) of RTOCs was determined against postharvest spoilage fungi of the genera Botrytis, Penicillium, Alternaria, and Monilinia, by measuring the reduction of the fungal biomass after exposure for 72 h at 25 °C. Thymol showed the lowest MIC (7.0 µg/L), followed by γ-terpinene (28.4 µg/L) and p-cymene (40.0 µg/L). In the case of P. digitatum ITEM 9569, resistant to commercial RTO, a better evaluation of interactions among RTOCs was performed using the checkerboard assay and the calculation of the Fractional Inhibitory Concentration Index (FICI). During incubation, changes in the RTOCs concentration were measured by GC-MS analysis. A synergistic effect between thymol (0.013 ± 0.003 L/L) and γ-terpinene (0.990 ± 0.030 L/L) (FICI = 0.50) in binary combinations, and between p-cymene (0.700 ± 0.010 L/L) and γ-terpinene (0.290 ± 0.010 L/L) in presence of thymol (0.008 ± 0.001 L/L) (FICI = 0.19), in ternary combinations was found. The synergistic effect against the strain P. digitatum ITEM 9569 suggests that different combinations among RTOCs could be defined to control fungal strains causing different food spoilage phenomena.

Combined application of antagonistic Wickerhamomyces anomalus BS91 strain and Cynara cardunculus L. leaf extracts for the control of postharvest decay of citrus fruit.

Combined use of biocontrol agents and plant extracts can be considered a viable and promising strategy for protecting plant tissues with different synergistic mechanisms of action that improve the antimicrobial activity of the mixtures. Treatments of citrus fruits with Wickerhamomyces anomalus BS91 have been previously reported as effective measures to reduce the incidence of green mold caused by Penicillium digitatum. On the opposite, the knowledge of the antifungal activity of cultivated cardoon (Cynara cardunculus L. var. altilis DC.) leaf extract, vegetable widespread in some Mediterranean areas, is still very limited. In this study, experimental trials were conducted to evaluate the effectiveness in vitro of leaf aqueous, methanolic and ethanolic extracts of C. cardunculus against seven fungal pathogens responsible for considerable food losses in the postharvest stage. In addition, biocontrol yeast W. anomalus BS91 and the three C. cardunculus extracts were tested in vivo both as a single treatment and in mixture, against Penicillium digitatum on 'Tarocco' oranges and 'Femminello' lemons. The combination of W. anomalus BS91 and leaf ethanolic extract reduced with the highest efficacy the incidence and severity of green mold on orange and lemon fruits with respect to the control, and was more effective than treatment with antagonistic yeast or leaf extracts applied alone. Incidence and severity of citrus decay were more consistently reduced when mixtures were applied 24 h before the inoculation of the pathogen, thus suggesting the relevance of preventive treatments. The mixtures of antagonistic W. anomalus BS91 and ethanolic leaf extract were more effective in controlling green mold decay on oranges than on lemons. These results indicate that biocontrol agents and leaf extracts, used in appropriate combination, can provide a stronger protection than when used singularly. However, compatibility between microbial antagonist and antimicrobial extract should be preliminary verified.

Effects of mung bean starch/guar gum-based edible emulsion coatings on the staling and safety of rice cakes.

Antimicrobial starch/gum-based edible emulsion coatings were developed to improve the storage stability of rice cakes by retarding starch retrogradation and inhibiting microbial growth. Rice cakes were coated with mung bean starch (MBS) and guar gum (GG) containing various concentrations of sunflower seed oil (SO). Among these, the (2 g MBS +0.75 g GG +1.5 g SO) / 100 g (optimum) decreased the hardness of rice cakes by 29 % and the crystallization rate (k) by 24 % compared with those of uncoated samples. The moisture loss in uncoated samples was markedly higher than that in the optimum blend-coated samples. Crystallinity analysis revealed the retarding effect of the developed coatings in starch retrogradation. Furthermore, adding 0.8 % (w/w) grapefruit seed extract to the optimum blend led to a distinct antimicrobial activity. Therefore, the newly developed edible coating was effective in maintaining the quality and safety of rice cakes.

Characterization and antifungal activity of jackfruit (Artocarpus heterophyllus Lam.) leaf extract obtained using conventional and emerging technologies.

With the current agro-food industry demands for more environmentally-friendly production, the use of natural antifungal compounds extracted by emerging technologies led to a sustainable alternative to control microorganisms. Therefore, the present work aimed to: (i) determine antifungal activity against Colletotrichum gloeosporioides and Penicillium italicum of jackfruit leaf extracts obtained by different methods, and (ii) identify compounds by HPLC-DAD-ESI-MS. Microwave (MAE), high-hydrostatic pressure (HHP) and ultrasound (UAE) assisted extractions were tested, followed by fractionation with different polarity solvents. The concentration of total soluble phenols (TSP), tannins (TT) and flavonoids (TF) were determined. Differences not only in the amounts of extracted phytochemicals were found but in the antifungal properties (MAE against P. italicum and HHP against C. gloeosporioides at 5 mg/ml) as well as in the chemical composition determined by HPLC-MS. These results suggest the possible application of jackfruit leaf extracts as a suitable postharvest antifungal treatment.

Development of Biodegradable Whey-Based Laminate Functionalised by Chitosan-Natural Extract Formulations.

In this research, antimicrobial polysaccharide chitosan and natural extracts were used as surface coating of a plastic laminate with an integrated whey layer on the inside. The aim was to establish the biodegradable and active concept of packaging laminates. For this purpose, chitosan nanoparticles (CSNPs) with embedded rosemary or cinnamon extracts were synthesised and characterised. Additionally, a whey-based laminate was functionalised: i) chitosan macromolecular solution was applied as first layer and ii) cinnamon or rosemary extracts encapsulated in CSNPs were applied as upper layer (layer wise deposition). Such functionalised whey-based laminate was physicochemically characterized in terms of elemental surface composition, wettability, morphology and oxygen permeability. The antimicrobial activity was tested against Staphylococcus aureus, Escherichia coli, Aspergillus flavus and Penicillium verrucosum. The antioxidant properties were determined using the ABTS assay. It could be shown that after functionalization of the films with the above-mentioned strategy, the wettability was improved. Furthermore, such whey-based laminates still show excellent barrier properties, good antimicrobial activity and a remarkable antioxidative activity. In addition to the improved biodegradability, this type of lamination could also have a positive effect on the shelf-life of products packaged in such structured films.

Assessment of Mint, Basil, and Lavender Essential Oil Vapor-Phase in Antifungal Protection and Lemon Fruit Quality.

There is an increasing interest in developing natural methods to replace the current chemicals used for maintaining postharvest quality of citrus fruits. The essential oil antifungal activity of mint (MEO), basil (BEO), and lavender (LEO) acting as the vapor-phases was tested against Penicillium digitatum. The minimum doses with fungistatic and fungicidal effect, in vitro, acting as the vapor-phases, were set up. The minimum fungicidal dose was 300 µL for BEO and 350 µL LEO, while for MEO only minimal dose with fungistatic effect was reached. The IC50 values were calculated and used (v/v) for testing preservation of lemon fruits, in close space enriched in vapor oil. For this purpose, the following two independent in vivo experiments were carried out: experiment 1, inoculated lemons with P. digitatum stored without chemical treatments 7 days, at 22 ± 2 °C, at two concentrations (C1-IC50 equivalent; C2-half of C1); and experiment 2, the non-inoculated lemons kept under the same conditions and concentrations of EO vapor served to evaluate the lemon quality properties. The results showed that antifungal protective effect was provided in the order of LEO-C1 > BEO-C1 > MEO-C1 > BEO-C2 > MEO-C2 > LEO-C2. The quality indicators like weight loss, pH, and firmness were not negatively influenced.

Effectiveness of essential oil extracted from pompia leaves against Penicillium digitatum.

BACKGROUND: In recent years, interest in the use of natural compounds as possible substitutes for chemicals, to prevent microbial food spoilage has grown. The antimicrobial activity of the essential oils (EOs) is well known and nowadays there is renewed interest in their application as natural preservatives in postharvest management. The aims of this study were to characterize the EO extracted from pompia leaves and to evaluate its effectiveness for the control of the postharvest decay agent Penicillium digitatum, when applied as vapor contact in new airtight boxes, supplied with a heating system. RESULTS: Fumigation was performed in vitro and on food using two concentrations of the EO, heated at controlled temperature. The headspace analysis revealed that the heating of the EO favored the evaporation of the volatile compounds, without altering their functionality. The treatments reduced the pathogen growth in vitro and rot on inoculated food by about 50%. CONCLUSION: The chemical analysis of the vapor composition demonstrated that heating the oil did not alter the components and thus the antimicrobial effect of the oil. The treatment by vapor contact with the EO was effective in controlling the pathogen growth in vitro but, above all, it was successful in halving rot in vivo. Due to their bioactivity in the vapor phase, EOs could be delivered as fumigants during postharvest protection; however, the techniques commonly employed are not ideal for simulating real pre-treatment conditions. The new device allows real large-scale conditions to be reproduced. © 2020 Society of Chemical Industry.

Green synthesis of garlic oil nanoemulsion using ultrasonication technique and its mechanism of antifungal action against Penicillium italicum.

Penicillium italicum (P. italicum) can cause significant economic loss of fruits and vegetables. Although garlic oil (GO) is an effective antimicrobial agent, the unstability and hydrophobicity limit its use as an environmentally friendly alternative to the conventional antibiotics against P. italicum. In this study, we focused on the fabrication and characterization of a functional GO nanoemulsion (NE) using ultrasonic technique and revealed the antifungal mechanism of the GO NE on P. italicum based on morphological, structural and molecular analyses. The optimal hydrophilic lipophilic balance (HLB) value determined for GO was 14 through the combination of Tween 80 and Span 80. Then the Box-Benhnken Design (BBD) was applied to produce the GO NE and the effects of different fabrication parameters on the particle size were evaluated. The optimal GO NE was selected with the GO concentration of 5.5%, the Smix concentration of 10%, the ultrasonic time of 5 min and the power of 50%. This GO NE had the smallest particle size of 52.27 nm, the best antifungal effect and the most stability. Furthermore, the antifungal mechanism of the GO NE on P. italicum was evaluated by extracellular conductivity, micro-Raman spectra, fluorescence imaging and scanning electron microscopy (SEM) imaging. The results presented that the GO NE retained the antifungal active ingredients. The fungal cell structure and morphology were malformed after treated with the GO NE and the lipids, nucleic acids and protein of P. italicum were destructed. Finally, the optimal GO NE was applied in vivo and P. italicum in citrus was successfully inhibited. It indicated that the optimal GO NE had the better antifungal activity against P. italicum than the pure GO. Besides, the minimum inhibitory concentration (MIC) of GO after preparing into the NE was changed from 3.7% to 0.01265% with about 300 times improvement of bioavailability. Therefore, the synthetic GO NE which promoted the bioavailability of GO was recommended as a promising alternative to inhibit P. italicum in vegetables and fruits.

Resistant fungi isolated from contaminated uranium mine in Brazil shows a high capacity to uptake uranium from water.

The Osamu Utsumi uranium mine occupies a 20 km2 area in the city of Caldas, which is located in the state of Minas Gerais, Brazil. Since mining activities ended at Osamu Utsumi 24 years ago, the surrounding area has become contaminated by acid effluents containing high concentrations of uranium. Thus, the aim of this study was to assess the uranium bioremediation capacity of 57 fungi isolated from the mine area. In tolerance tests, 38% (22) of the fungal isolates were considered tolerant to uranium, including 10 Penicillium species. At a uranium concentration of 2000 mg L-1 48 fungi did not exhibit mycelial growth index inhibition. Minimal inhibitory concentration (MIC) analysis showed growth of 25 fungi above a uranium concentration of 8000 mg L-1. At high uranium concentrations, some fungi (i.e., Talaromyces amestolkiae and Penicillium citrinum) showed morphological changes and pigment (melanin) production. Among the fungal isolates, those considered to be more tolerant to uranium were isolated from soil and sediment samples containing higher concentrations of heavy metal. When comparing the results of resistance/tolerance tests with those for uranium biosorption capacity, we concluded that the fungi isolated from the Osamu Utsumi mine with the best potential for uranium bioremediation were Gongronella butleri, Penicillium piscarium, Penicillium citrinum, Penicillium ludwigii, and Talaromyces amestolkiae. Biosorption tests with live fungal biomass showed that 11 species had a high potential for uranium uptake from contaminated water.

Quality Control Standardization and Evaluation of Antimicrobial Potential of Daruhaldi (Berberis aristata DC) Stem Bark.

Berberis aristata is used for the treatment of diabetes, piles, and liver diseases. As the drug is broadly used in Indigenous systems of medicine, it was designed to set the quality standards and antimicrobial potential for the stem bark of Berberis aristata. Botanical, physicochemical, pharmacotoxicological, fluorescence, microbial load, and phytochemical parameters of the stem bark were determined. High-performance thin-layer chromatography (HPTLC) was carried out by the CAMAG-HPTLC system. Berberine, total phenolics, and flavonoids were estimated. The antimicrobial potential was determined against the bacteria Bacillus subtilis and Escherichia coli and fungi Penicillium citrinum and Aspergillus terreus. The foreign matter, foaming index, swelling index, bitterness value, resin content, loss on drying, total ash, acid-insoluble ash, water-soluble ash, heavy metals, microbial load, berberine content, total phenolic content, and total flavonoid content were found to be 0, 0, 5, 1.34, 0.86%, 2.07%, 4.33%, 0.28%, 2.66%, within limits, 6 colonies in 1/100 dilution, 0.032 mg/g, 144.04 µg/ml, and 85.61 µg/ml, respectively. Phytochemicals such as phenolics, flavonoids, and sterols were present in the methanolic extract. The fluorescences observed in UV light were of different colors in different solvents. The methanolic extract and standards exhibited antimicrobial activity at the tested concentrations against the microbial strains. Results confirmed the quality and purity of the drug B. aristata. Results also confirmed that methanolic extract of B. aristata stem bark possesses potent antimicrobial activity. Thus, the use of this quality-controlled plant-derived drug with established antimicrobial property could be of great significance in quality-control standardization and preventive and therapeutic approaches to infectious diseases.

Ammoides verticillata Essential Oil as Biocontrol Agent of Selected Fungi and Pest of Olive Tree.

BACKGROUND: Ammoides verticillata Briq is an aromatic and medicinal herb that has been widely used in folk medicine for treatment of several types of pathologies such as respiratory problems, colds, fever, headache, gastric disorders and renal infections. OBJECTIVE: Essential oils have been subjected of numerous works for their antimicrobial and insecticidal properties which have been valued by hundreds of publications and patents. The main objective of this work was to evaluate the chemical composition, and the in vitro and in vivo antifungal, and insecticidal properties of essential oil of Ammoides verticillata. METHODS: In this work, the air-dried aerial parts of Ammoides verticillata were hydrodistilled in a Clevenger-type apparatus. The essential oil isolated was analyzed using gas chromatography (GC) and mass spectrometry (GC/MS). The in vitro antifungal activity of the essential oil was investigated against four plant fungi using radial growth technique. The effect of the essential oil on disease development of olive caused by Penicillium crustosum and Aspergillus niger in the in vivo conditions. The fumigation test of the essential oil was evaluated against L3 stages, pupas and adults of Bactrocera oleae known as the olive fly. RESULTS: The essential oil of A. verticillata was characterized mainly by carvacrol (44.3%), limonene (19.3%) and p-cymene (19.2%). The result of in vitro antifungal activity of essential oil showed an interesting antifungal inhibition against Alternaria alternata and Fusarium solani strains with percentage inhibition of 89%. Furthermore, oil of A. verticillata has demonstrated promising in vivo antifungal activity to control infections of olives caused by Aspergillus niger and Penicillium crustosum. In addition, the essential oil exhibited fumigation toxicity against Bactrocera oleae with mortality percentage of 100% at 2 µL/L air. CONCLUSION: The results showed that essential oil of A. verticillata has interesting biological activities, which suggests that oil have the potential to be used as biocide to protect olives of pathogenic fungi and pests.

Investigation of the antifungal effects of algal extracts on apple-infecting fungi.

It is known that cell extracts of various algae have antifungal activity against microorganisms in vitro. Antifungal activities of Ulva lactuca, Chlorella vulgaris, Chlorella minutissima, and Chlorella protothecoides were investigated against: Aspergillus niger, Alternaria alternata, and Penicillium expansum fungi to present their fungicide potentials. Aspergillus niger, Alternaria sp., and Penicillium expansum are typical soft-rotting fungi and cause important loss of apple fruit in the storage. In vitro antifungal activity was evaluated by agar disc diffusion assay against pathogenic apple rot fungi. As a result, almost all of the extracts obtained from algae species were revealed to have antifungal activity against selected fungal pathogens. Free radical-scavenging activity of the extracts was determined with 1,1-diphenyl-2 picryl hydrazyl (DPPH) free radical-scavenging method. Extract of C. protothecoides was determined to have a stronger antioxidant activity than other algae extracts. This study reveals that the potential of algae should be investigated for the production of natural fungicide for pharmaceutical and food industries.

Cold Plasma Treatment as an Alternative for Ochratoxin a Detoxification and Inhibition of Mycotoxigenic Fungi in Roasted Coffee.

Ochratoxin A (OTA) produced by mycotoxigenic fungi (Aspergillus and Penicillium spp.) is an extremely toxic and carcinogenic metabolite. The use of cold plasma to inhibit toxin-producing microorganisms in coffee could be an important alternative to avoid proliferation of mycotoxigenic fungi. Roasted coffee samples were artificially inoculated with A. westerdijikiae, A. steynii, A. versicolor, and A. niger, and incubated at 27 °C over 21 days for OTA production. Samples were cold plasma treated at 30 W input power and 850 V output voltage with helium at 1.5 L/min flow. OTA production in coffee was analyzed by high performance liquid chromatography coupled to a mass spectrometer (HPLC-MS). After 6 min of treatment with cold plasma, fungi were completely inhibited (4 log reduction). Cold plasma reduces 50% of OTA content after 30 min of treatment. Toxicity was estimated for extracts of artificially contaminated roasted coffee samples using the brine shrimp (Artemia salina) lethality assay. Toxicity for untreated roasted coffee was shown to be "toxic", while toxicity for cold plasma treated coffee was reduced to "slightly toxic". These results suggested that cold plasma may be considered as an alternative method for the degradation and reduction of toxin production by mycotoxigenic fungi in the processing of foods and feedstuffs.