Antifungal Activities of Biogenic Silver Nanoparticles Mediated by Marine Algae: In Vitro and In Vivo Insights of Coating Tomato Fruit to Protect against Penicillium italicum Blue Mold.

In an attempt to reduce such decay induced by pathogenic causes, several studies investigated the effectiveness of nanoparticles (NPs) that play a vital role in saving food products, especially fruits. Current research delves into biogenic silver nanoparticles (using marine alga Turbinaria turbinata (Tt/Ag-NPs) and their characterization using FT-IR, TEM, EDS, and zeta potential. Some pathogenic fungi, which cause fruit spoilage, were isolated. We studied the impact of using Tt/Ag-NPs to protect against isolated fungi in vitro, and the influence of Tt/Ag-NPs as a coating of tomato fruit to protect against blue mold caused by Penicillium italicum (OR770486) over 17 days of storage time. Five treatments were examined: T1, healthy fruits were used as the positive control; T2, healthy fruits sprayed with Tt/Ag-NPs; T3, fruits infected with P. italicum followed by coating with Tt/Ag-NPs (pre-coating); T4, fruits coated with Tt/Ag-NPs followed by infection by P. italicum (post-coating); and T5, the negative control, fruits infected by P. italicum. The results displayed that Tt/Ag-NPs are crystalline, spherical in shape, with size ranges between 14.5 and 39.85 nm, and negative charges. Different concentrations of Tt/Ag-NPs possessed antifungal activities against Botrytis cinerea, Rhodotorula mucilaginosa, Penicillium expansum, Alternaria alternate, and Stemphylium vesicarium. After two days of tomatoes being infected with P. italicum, 55% of the fruits were spoilage. The tomato fruit coated with Tt/Ag-NPs delayed weight loss, increased titratable acidity (TA%), antioxidant%, and polyphenol contents, and decreased pH and total soluble solids (TSSs). There were no significant results between pre-coating and post-coating except in phenol contents increased in pre-coating. A particular focus is placed on the novel and promising approach of utilizing nanoparticles to combat foodborne pathogens and preserve commodities, with a spotlight on the application of nanoparticles in safeguarding tomatoes from decay.

Comparative physiological and transcriptomic analysis reveals an improved biological control efficacy of Sporidiobolus pararoseus Y16 enhanced with ascorbic acid against the oxidative stress tolerance caused by Penicillium expansum in pears.

Sporidiobolus pararoseus Y16, a species of significant ecological importance, has distinctive physiological and biological regulatory systems that aid in its survival and environmental adaptation. The goal of this investigation was to understand the complex interactions between physiological and molecular mechanisms in pear fruits as induced by S. pararoseus Y16. The study investigated the use of S. pararoseus Y16 and ascorbic acid (VC) in combination in controlling blue mold decay in pears via physiological and transcriptomic approach. The study results showed that treatment of S. pararoseus Y16 with 150 µg/mL VC reduced pears blue mold disease incidence from 43% to 11%. Furthermore, the combination of S. pararoseus Y16 and VC significantly inhibited mycelia growth and spore germination of Penicillium expansum in the pear's wounds. The pre-treatment did not impair post-harvest qualities of pear fruit but increased antioxidant enzyme activity specifically polyphenol oxidase (PPO), peroxidase (POD), catalase (CAT) activities as well as phenylalanine ammonia-lyase (PAL) enzyme activity. The transcriptome analysis further uncovered 395 differentially expressed genes (DEGs) and pathways involved in defense mechanisms and disease resistance. Notable pathways of the DEGs include plant-pathogen interaction, tyrosine metabolism, and hormone signal transduction pathways. The integrative approach with both physiological and transcriptomic tools to investigate postharvest pathology in pear fruits with clarification on how S. pararoseus Y16 enhanced with VC, improved gene expression for disease defense, and create alternative controls strategies for managing postharvest diseases.

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.

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.

Novel Chitosan-Riboflavin Conjugate with Visible Light-Enhanced Antifungal Properties against Penicillium digitatum.

A novel chemical conjugate between chitosan (CH) and riboflavin (RF) has been synthesized and characterized via Fourier transform infrared, NMR, and other spectroscopic methods. Photophysical and photochemical properties such as absorption spectra, fluorescence emission, fluorescence anisotropy, and singlet oxygen generation were characterized as well. This new biopolymer-based conjugate was designed to have an antifungal effect enhanced through antimicrobial photodynamic therapy. The antifungal effect of this conjugate (CH-RF) was compared with CH and RF against Penicillium digitatum in vitro. The conjugate showed the highest fungal growth inhibition of all systems tested at a dose of 0.5% w/v. This new biopolymer-based compound could be a promising alternative to fungicides used in citrus fruits postharvest.

Effect of red thyme oil (Thymus vulgaris L.) vapours on fungal decay, quality parameters and shelf-life of oranges during cold storage.

This work has been aimed at studying the effect of red thyme oil (RTO, Thymus vulgaris L.) on the shelf-life and Penicillium decay of oranges during cold storage. RTO vapours significantly reduced (P ≤ 0.05) the percentage of infected wounds, the external growth area and the production of spores in inoculated orange fruit stored for 12 days at 7 °C in a polypropylene film selected for its appropriate permeability. Among the RTO compounds, p-cymene and thymol were the most abundant in packed boxes at the end of cold storage. The RTO vapours did not affect the main quality parameters of the oranges, or the taste and odour of the juice. The results have shown that an active packaging, using RTO vapours, could be employed, by the citrus industry, to extend the shelf-life of oranges for fresh market use and juice processing.

Antifungal activity of an allicin derivative against Penicillium expansum via induction of oxidative stress.

The application of natural preservatives has become an attractive method for controlling postharvest decay of fruits and vegetables. Allicin, the main active ingredient of allium plants, has broad-spectrum antifungal activity. However, the unstable properties of allicin limit its wide application. In this study, 1-[(R)-ethylsulfinyl]sulfanylethane, a structurally stable derivative of allicin, was used to explore its antifungal activity and potential mechanism on the expansion of Penicillium expansum. We demonstrated the antifungal activity of 1-[(R)-ethylsulfinyl]sulfanylethane through in vitro antifungal experiments. At the concentration is 2 mg/L, 1-[((R)-ethylsulfinyl]sulfanyl]ethane can completely inhibit spore germination and mycelial growth, whereas the concentration of allicin needs to reach 16 mg/L. Fungal Biochemical assay indicated that decrease of mitochondrial membrane potential, overgeneration of reactive oxygen species, decrease of adenosine triphosphate and glutathione content, increase of superoxide dismutase, catalase, and malondialdehyde content. The results revealed that 1-[(R)-ethylsulfinyl]sulfanylethane induced mitochondrial dysfunction and oxidative stress in P. expansum. Due to its excellent antifungal activity, 1-[((R)-ethylsulfinyl]sulfanyl]ethane might be developed as a substitute for fungicides against P. expansum in postharvest fruit storage.

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.

Antioxidant and antibacterial activities, interfacial and emulsifying properties of the apo and holo forms of purified camel and bovine α-lactalbumin.

The antioxidant and antibacterial activities of camel and bovine α-lactalbumin (α-La) in both calcium-loaded (holo) and calcium-depleted (apo) forms were investigated and compared. Antioxidant assay showed that camel and bovine α-La exhibited significant Ferric-reducing antioxidant power (FRAP), ferrous iron-chelating activity (FCA) and antiradical activities especially in their apo form. Camel apo α-La also exhibited attractive antibacterial activities against Gram-negative bacteria (Pseudomonas aeruginosa) and against fungal pathogens species (Penicillium bilaiae, Aspergillus tamari and Aspergillus sclerotiorum). Likewise, emulsifying properties (emulsification ability (EAI) and stability (ESI) indexes) and the surface characteristics (surface hydrophobicity, ζ-potential and interfacial tension) of the α-La were assessed. Maximum EAI were found at pH 7.0, with higher EAI values for the camel apo α-La (EAI ~19.5 m2/g). This behavior was explained by its relative high surface hydrophobicity and its greater efficiency to reduce the surface tension at the oil-water interface. Furthermore, emulsions were found to be more stable at pH 7.0 compared to pH 5.0 (ESI ~50%) due to the higher electrostatic repulsive forces between oil droplets at pH 7.0 in consistence with the ζ-potential results. This study concluded that the camel apo α-La has antibacterial, antioxidant, and emulsifying properties in agricultural and food industries.

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.

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.

Ca2+ and SO42- accelerate the reduction of Cr(VI) by Penicillium oxalicum SL2.

Some ions in soils may affect the growth and metabolism of microorganisms and subsequently alter the remediation efficiency of Cr(VI). Here, the effects of different Ca2+ and SO42- levels on the reduction of Cr(VI) by Penicillium oxalicum SL2 were investigated. The results showed that Cr(VI) reduction by P. oxalicum SL2 in potato dextrose liquid (PDL) medium was accelerated by the presence of exogenous Ca2+ and SO42-. The Cr(VI) reduction rates were increased by 52.5% (200 mg L-1 Ca2+ treated) and 55.9% (2000 mg L-1 SO42- treated), respectively. High concentration of Ca2+ in medium resulted in the production of calcium oxalate crystals, which was contributed to the adsorption of chromium. In addition, X-ray absorption near-edge spectroscopy (XANES) analysis showed that P. oxalicum SL2 could reduce the toxicity of Cr(VI) by synthesizing cysteine (Cys) and reduced glutathione (GSH). The decrease of thiol compounds (Cys and GSH) in P. oxalicum SL2 mycelia treated with SO42- proved the alleviation of oxidative stress. In conclusion, exogenous Ca2+ could reduce the damage of Cr(VI) to P. oxalicum SL2 by maintaining the integrity of cell wall, and the addition of SO42- alleviated the Cr(VI) toxicity to P. oxalicum SL2, thus accelerating the reduction of Cr(VI).

A Case Series and Literature Review of Isavuconazole Use in Pediatric Patients with Hemato-oncologic Diseases and Hematopoietic Stem Cell Transplantation.

We analyzed the use of isavuconazole (ISA) as treatment or prophylaxis for invasive fungal disease (IFD) in children with hemato-oncologic diseases. A multicentric retrospective analysis was performed among centers belonging to the Italian Association for Pediatric Hematology and Oncology (AIEOP). Pharmacokinetic (PK) monitoring was applied by a high-performance liquid chromatography-tandem mass spectrometry (HLPC-MS/MS) assay. Twenty-nine patients were studied: 10 during chemotherapy and 19 after allogeneic hematopoietic stem cell transplantation (HSCT). The patients consisted of 20 males and 9 females with a median age of 14.5 years (age range, 3 to 18 years) and a median body weight of 47 kg (body weight range, 15 to 80 kg). ISA was used as prophylaxis in 5 patients and as treatment in 24 cases (20 after therapeutic failure, 4 as first-line therapy). According to European Organization for Research and Treatment of Cancer (EORTC) criteria, we registered 5 patients with proven IFD, 9 patients with probable IFD, and 10 patients with possible IFD. Patients with a body weight of <30 kg received half the ISA dose; the others received ISA on the adult schedule (a 200-mg loading dose every 8 h on days 1 and 2 and a 200-mg/day maintenance dose); for all but 10 patients, the route of administration switched from the intravenous route to the oral route during treatment. ISA was administered for a median of 75.5 days (range, 6 to 523 days). The overall response rate was 70.8%; 12 patients with IFD achieved complete remission, 5 achieved partial remission, 5 achieved progression, and 3 achieved stable IFD. No breakthrough infections were registered. PK monitoring of 17 patients revealed a median ISA steady-state trough concentration of 4.91 mg/liter (range, 2.15 to 8.54 mg/liter) and a concentration/dose (in kilograms) ratio of 1.13 (range, 0.47 to 3.42). Determination of the 12-h PK profile was performed in 6 cases. The median area under the concentration-time curve from 0 to 12 h was 153.16 mg·h/liter (range, 86.31 to 169.45 mg·h/liter). Common Terminology Criteria for Adverse Events grade 1 to 3 toxicity (increased transaminase and/or creatinine levels) was observed in 6 patients, with no drug-drug interactions being seen in patients receiving immunosuppressants. Isavuconazole may be useful and safe in children with hemato-oncologic diseases, even in the HSCT setting. Prospective studies are warranted.

Halogenated boroxine dipotassium trioxohydroxytetrafluorotriborate K2[B3O3F4OH] inhibits emerging multidrug-resistant and ß-lactamase-producing opportunistic pathogens.

Halogenated boroxine dipotassium trioxohydroxytetrafluorotriborate, K2[B3O3F4OH] (boroxine) was previously shown to be very effective in inhibition of several carcinoma cell lines, including the skin cancer. Here, we investigated its antimicrobial potential by targeting the multidrug-resistant opportunistic pathogens associated with skin and wound infections. The antimicrobial testing against eleven bacterial and four fungal species revealed good activity of boroxine against pathogenic filamentous fungi Penicillium funiculosum and Aspergillus niger (MIC50 64 and 128 µg/ml), and a moderate bioactivity against the yeast Candida albicans (MIC50 512 µg/ml). Among the tested multidrug-resistant bacteria, the best antibacterial effect, stable over a 24-h period, was observed against the methicillin-resistant Staphylococcus aureus strain (MRSA) at MIC of 1024 µg/ml. The atomic force microscopy (AFM) used to investigate the morphology of S. aureus cells revealed indentations on its cell envelope after the boroxine exposure. These results show that in addition to the antitumor effect, boroxine exerts wide spectrum antimicrobial activity, thus may help preventing the development of skin and wound-related opportunistic infections.

Development of a Microbial Decontamination System Combining Washing with Highly Activated Calcium Oxide Solution and Antimicrobial Coating for Improvement of Mandarin Storability.

A new microbial decontamination system combining washing with a natural antimicrobial solution and coating with a carnauba wax (CW)-based antimicrobial coating was developed and its effects on mandarin storability were investigated. Mandarins were washed with an antimicrobial solution and/or coated with grapefruit seed extract-CW (GSE/CW). Values for the disease incidence of Penicillium digitatum in untreated mandarins; mandarins coated with GSE/CW without washing; and mandarins coated with GSE/CW after washing with a fumaric acid (FA) solution of slightly acidic electrolyzed water, a highly activated calcium oxide (CaO) aqueous solution, or CaO solution followed by FA solution were 96.0, 70.0, 78.8, 50.0, and 72.2%, respectively. GSE/CW coating after CaO washing was most effective in inhibiting P. digitatum growth during storage at 25 °C. Compared to untreated samples, GSE/CW coating alone or after CaO washing retained CO2 generation, firmness, and total polyphenol content of mandarins at 25 °C. Such treatments also effectively maintained mandarin pH, ascorbic acid concentration, and antioxidant capacity at both 4 and 25 °C. Moreover, GSE/CW coating after CaO washing more effectively inhibited P. digitatum growth at 25 °C and maintained ascorbic acid concentration and antioxidant capacity at 4 and 25 °C than GSE/CW coating alone. The microbial decontamination system integrating CaO washing and GSE/CW coating demonstrates potential for improving mandarin storability by inhibiting P. digitatum growth and improving the preservation of quality properties and sensory characteristics. PRACTICAL APPLICATION: This is the first study to develop a microbial decontamination system involving both washing with a natural antimicrobial solution and carnauba wax coating containing grapefruit seed extract to improve the storability of fruits. This system demonstrated a primary effect of inhibiting fungi that cause mandarin surface decay at 25 °C via the highly activated calcium oxide wash and secondary effects of delaying quality degradation and inhibiting fungal growth by the action of the antimicrobial coating. These effects led to improvements in mandarin storability, along with enhanced visual appeal while not affecting taste, flavor, or texture.

The multi-layer film system improved the release and retention properties of cinnamon essential oil and its application as coating in inhibition to penicillium expansion of apple fruit.

The aim of this research is to develop, characterize and utilize a multi-layer antibacterial film using chitosan (CS) and sodium alginate (SA) as biopolymers and cinnamon essential oil (CEO) as main antibacterial ingredients. The dense cross-section of SA layer in the scanning electron microscopy (SEM) analysis verified that layer-by-layer method improved physical and mechanical properties of CS-CEO single layer film. The thermogravimetric (TGA) and fourier transform infrared (FT-IR) analysis indicated that the layer-by-layer method changed the intermolecular interaction and the thermal stability. Importantly, the multi-layer film exhibited more sustained release and higher retention rate of CEO compared CS-CEO single layer film. The multi-layer coating showed a more significant and lasting inhibition of penicillium expansion which further demonstrated that the layer-by-layer method improved the release and retention of CEO in the multiphased system. To summarize, the multilayer film system is a promising controllable release system for loading essential oils.

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.