Optical Nanomotion Detection to Rapidly Discriminate between Fungicidal and Fungistatic Effects of Antifungals on Single-Cell Candida albicans.

Candida albicans is an emerging pathogen that poses a significant challenge due to its multidrug-resistant nature. There are two types of antifungal agents, fungicidal and fungistatic, with distinct mechanisms of action against fungal pathogens. Fungicidal agents kill fungal pathogens, whereas fungistatic agents inhibit their growth. The growth can be restored once the agent is removed and favorable conditions are established. Recognizing this difference is crucial as it influences treatment selection and infection prognosis. We present a technique based on optical nanomotion detection (ONMD) (i.e., observing the movement of the cells using an optical microscope) to discriminate rapidly between fungicidal (caspofungin) and fungistatic (fluconazole) drugs. The technique is based on the change in a yeast cell's nanomotion as a function of time during a two-hour treatment with the antifungal of interest followed by a one-hour growth period. The cells are entrapped in microwells in a microfluidic chip, which allows a quick exchange of growth medium and antifungal agent, enabling ONMD measurements on the same individual cells before and after treatment. This procedure permits to discriminate between fungicidal and fungistatic antifungals in less than 3 h, with single-cell resolution by observing if the nanomotion recovers after removing the treatment and reintroducing growth medium (YPD), or continues to drop. The simplicity of the approach holds promise for further development into a user-friendly device for rapid antifungal susceptibility testing (AFST), potentially being implemented in hospitals and medical centers worldwide in developed and developing countries.

Antimicrobial, antioxidant and antiproliferative activities of a galactose-binding seed lectin from Manilkara zapota.

A seed lectin from Manilkara zapota (MZSL) was purified using ammonium sulphate precipitation and affinity chromatography. Hemagglutination activity, neutral sugar content and physicochemical properties of the lectin were determined and toxicity was checked by brine shrimp toxicity assay. Antimicrobial, antioxidant as well as in vitro anticancer activities of MZSL were also evaluated. Our findings showed the molecular weight of MZSL to be 33.0 ± 1 kDa. Minimum hemagglutination concentration of the lectin was 15.625 µg/ml. With a neutral sugar content of 6.32 %, the lectin was fully active at a temperature range of 30-50 °C and pH 7.0-8.0 and it was mildly toxic with an LC50 value of 107.93 µg/ml. The lectin demonstrated bacteriostatic activity against gram-positive bacteria in contrast to gram-negative bacteria at a concentration of 31.25 µg/ml, agglutinated Staphylococcus aureus and Shigella dysenteriae and exerted fungistatic activity against Aspergillus niger. MZSL dose-dependently reduced the formation of biofilm by E. coli. DPPH assay confirmed its antioxidant activity with an IC50 value of 96.42 µg/ml. MZSL showed 21.64 % growth inhibition against Ehrlich ascites carcinoma (EAC) cells at 80 µg/ml whereas its antiproliferative potential against MCF-7 and A-549 cancer cell lines became evident with IC50 values of 70.66 µg/ml and 107.64 µg/ml, respectively.

Antimicrobial Properties of New Polyamines Conjugated with Oxygen-Containing Aromatic Functional Groups.

Antibiotic resistance is now a first-order health problem, which makes the development of new families of antimicrobials imperative. These compounds should ideally be inexpensive, readily available, highly active, and non-toxic. Here, we present the results of our investigation regarding the antimicrobial activity of a series of natural and synthetic polyamines with different architectures (linear, tripodal, and macrocyclic) and their derivatives with the oxygen-containing aromatic functional groups 1,3-benzodioxol, ortho/para phenol, or 2,3-dihydrobenzofuran. The new compounds were prepared through an inexpensive process, and their activity was tested against selected strains of yeast, as well as Gram-positive and Gram-negative bacteria. In all cases, the conjugated derivatives showed antimicrobial activity higher than the unsubstituted polyamines. Several factors, such as the overall charge at physiological pH, lipophilicity, and the topology of the polyamine scaffold were relevant to their activity. The nature of the lipophilic moiety was also a determinant of human cell toxicity. The lead compounds were found to be bactericidal and fungistatic, and they were synergic with the commercial antifungals fluconazole, cycloheximide, and amphotericin B against the yeast strains tested.

The performance of plant essential oils against lactic acid bacteria and adverse microorganisms in silage production.

Plant essential oils have played an important role in the field of antibiotic alternatives because of their efficient bacteriostatic and fungistatic activity. As plant essential oils are widely used, their activity to improve the quality of plant silage has also been explored. This review expounds on the active ingredients of essential oils, their bacteriostatic and fungistatic activity, and mechanisms, as well as discusses the application of plant essential oils in plant silage fermentation, to provide a reference for the development and application of plant essential oils as silage additives in plant silage fermentation feed.

Biological and Chemical Characterization of Musa paradisiaca Leachate.

There is a growing demand for molecules of natural origin for biocontrol and biostimulation, given the current trend away from synthetic chemical products. Leachates extracted from plantain stems were obtained after biodegradation of the plant material. To characterize the leachate, quantitative determinations of nitrogen, carbon, phosphorus, and cations (K+, Ca2+, Mg2+, Na+), Q2/4, Q2/6, and Q4/6 absorbance ratios, and metabolomic analysis were carried out. The potential role of plantain leachates as fungicide, elicitor of plant defense, and/or plant biostimulant was evaluated by agar well diffusion method, phenotypic, molecular, and imaging approaches. The plant extracts induced a slight inhibition of fungal growth of an aggressive strain of Colletotrichum gloeosporioides, which causes anthracnose. Organic compounds such as cinnamic, ellagic, quinic, and fulvic acids and indole alkaloid such as ellipticine, along with some minerals such as potassium, calcium, and phosphorus, may be responsible for the inhibition of fungal growth. In addition, jasmonic, benzoic, and salicylic acids, which are known to play a role in plant defense and as biostimulants in tomato, were detected in leachate extract. Indeed, foliar application of banana leachate induced overexpression of LOXD, PPOD, and Worky70-80 genes, which are involved in phenylpropanoid metabolism, jasmonic acid biosynthesis, and salicylic acid metabolism, respectively. Leachate also activated root growth in tomato seedlings. However, the main impact of the leachate was observed on mature plants, where it caused a reduction in leaf area and fresh weight, the remodeling of stem cell wall glycopolymers, and an increase in the expression of proline dehydrogenase.

Postharvest Diseases of Pomegranate: Alternative Control Means and a Spiderweb Effect.

The pomegranate is a fruit known since ancient times for its beneficial properties. It has recently aroused great interest in the industry and among consumers, leading to a significant increase in demand. Consequently, its cultivation has been boosted all over the world. The pomegranate crop suffers considerable yield losses, especially at the postharvest stage, because it is a "minor crop" with few permitted control means. To control latent (Alternaria spp., Botrytis spp., Coniella spp., Colletotrichum spp., and Cytospora spp.) and wound (Aspergillus spp., Penicillium spp., and Talaromyces spp.) fungal pathogens, different alternative compounds, previously evaluated in vitro, were tested in the field on pomegranate cv. Wonderful. A chitosan solution, a plant protein hydrolysate, and a red seaweed extract were compared with a chemical control treatment, all as preharvest (field application) and postharvest treatments and their combinations. At the end of the storage period, the incidence of stamen infections and external and internal rots, and the severity of internal decay were evaluated. Obtained data revealed that pre- and postharvest application of all substances reduced the epiphytic population on stamens. Preharvest applications of seaweed extract and plant hydrolysate were the most effective treatments to reduce the severity of internal pomegranate decays. Furthermore, the influence of spider (Cheiracanthium mildei) cocoons on the fruit calyx as a possible barrier against postharvest fungal pathogens was assessed in a 'Mollar de Elche' pomegranate organic orchard. Compared to no-cocoon fruit (control), the incidence of infected stamens and internal molds in those with spiderwebs was reduced by about 30%, and the mean severity of internal rots was halved. Spiderwebs analyzed via Scanning Electron Microscopy (SEM) disclosed a layered, unordered structure that did not allow for the passage of fungal spores due to its mean mesh size (1 to 20 µm ca). The aims of this research were (i) to evaluate alternative compounds useful to control postharvest pomegranate decays and (ii) to evaluate the effectiveness of spiders in reducing postharvest fungal infections by analyzing related mechanisms of action. Alternative control means proposed in the present work and calyx spider colonization may be helpful to reduce postharvest pomegranate diseases, yield losses, and waste production in an integrated control strategy, satisfying organic agriculture and the planned goals of Zero Hunger Challenge launched by the United Nations.

Limosilactobacillus reuteri as sustainable biological control agent against toxigenic Fusarium verticillioides.

Corn contamination with Fusarium verticillioides (Sacc.) Nirenberg is a worldwide problem that affects yield and grain quality resulting in severe economic losses and implications for food safety. Control of F. verticillioides is a challenge, but lactic acid bacteria (LAB) has high potential as a biological control agent. In this study, the antifungal effect of Limosilactobacillus reuteri (formerly Lactobacillus reuteri) LR-92 against F. verticillioides 97L was investigated. Cell-free supernatant (CFS) from L. reuteri showed concentration-dependent fungicidal and fungistatic activity against F. verticillioides 97L. The antifungal compounds from CFS showed heat stability and pH dependence, and antifungal activity was not affected by treatment with proteolytic enzymes. High-performance liquid chromatography analysis indicated that L. reuteri LR-92 produces lactic and acetic acids. After liquid-liquid extraction, electrospray ionization mass spectrometry analysis of the active ethyl acetate fraction containing antifungal compounds revealed the production of 3-phenyllactic acid, cyclo-(L-Pro-L-Leu), cyclo-(L-Pro-L-Phe), and cyclo-(L-Phe-trans-4-OH-L-Pro). L. reuteri LR-92 has potential as a biocontrol agent for F. verticillioides and contributes to food safety.

Chemo-enzymatic synthesis and biological activity evaluation of propenylbenzene derivatives.

Propenylbenzenes, including isosafrole, anethole, isoeugenol, and their derivatives, are natural compounds found in essential oils from various plants. Compounds of this group are important and valuable, and are used in the flavour and fragrance industries as well as the pharmaceutical and cosmetic industries. The aim of this study was to develop an efficient process for synthesising oxygenated derivatives of these compounds and evaluate their potential biological activities. In this paper, we propose a two-step chemo-enzymatic method. The first step involves the synthesis of corresponding diols 1b-5b from propenylbenzenes 1a-5avia lipase catalysed epoxidation followed by epoxide hydrolysis. The second step involves the microbial oxidation of a diasteroisomeric mixture of diols 1b-5b to yield the corresponding hydroxy ketones 1c-4c, which in this study was performed on a preparative scale using Dietzia sp. DSM44016, Rhodococcus erythropolis DSM44534, R. erythropolis PCM2150, and Rhodococcus ruber PCM2166. Application of scaled-up processes allowed to obtain hydroxy ketones 1-4c with the following yield range 36-62.5%. The propenylbenzene derivatives thus obtained and the starting compounds were tested for various biological activities, including antimicrobial, antioxidant, haemolytic, and anticancer activities, and their impact on membrane fluidity. Fungistatic activity assay against selected strains of Candida albicans results in MIC50 value varied from 37 to 124 µg/mL for compounds 1a, 3a-c, 4a,b, and 5a,b. The highest antiradical activity was shown by propenylbenzenes 1-5a with a double bond in their structure with EC50 value ranged from 19 to 31 µg/mL. Haemolytic activity assay showed no cytotoxicity of the tested compounds on human RBCs whereas, compounds 2b-4b and 2c-4c affected the fluidity of the RBCs membrane. The tested compounds depending on their concentration showed different antiproliferative activity against HepG2, Caco-2, and MG63. The results indicate the potential utility of these compounds as fungistatics, antioxidants, and proliferation inhibitors of selected cell lines.

Characterization and Inhibitory Effects of Essential Oil and Nanoemulsion from Ocotea indecora (Shott) Mez in Aspergillus Species.

The Aspergillus genus, the etiological agent of aspergillosis, is an important food contaminant and mycotoxin producer. Plant extracts and essential oils are a source of bioactive substances with antimicrobial potential that can be used instead of synthetic food preservatives. Species from the Lauraceae family and the Ocotea genus have been used as traditional medicinal herbs. Their essential oils can be nanoemulsified to enhance their stability and bioavailability and increase their use. Therefore, this study sought to prepare and characterize both nanoemulsion and essential oil from the Ocotea indecora's leaves, a native and endemic species from the Mata Atlântica forest in Brazil, and evaluate the activity against Aspergillus flavus RC 2054, Aspergillus parasiticus NRRL 2999, and Aspergillus westerdjikiae NRRL 3174. The products were added to Sabouraud Dextrose Agar at concentrations of 256, 512, 1024, 2048, and 4096 µg/mL. The strains were inoculated and incubated for up to 96 h with two daily measurements. The results did not show fungicidal activity under these conditions. A fungistatic effect, however, was observed. The nanoemulsion decreased the fungistatic concentration of the essential oil more than ten times, mainly in A. westerdjikiae. There were no significant changes in aflatoxin production.

Withania somnifera seed oil exhibits antibiofilm properties against drug-resistant Candida auris clinical isolate through modulation in cell permeability.

AIM: Candida auris, fast evolving drug-resistant fungus, poses an imminent global health threat. Alternative drug-resistance nonevoking treatment options are necessary. This study explored the antifungal and antibiofilm efficacies of Withania somnifera seed oil extracted using super critical CO2 (WSSO) against clinically isolated Fluconazole-resistant C. auris and its putative mode-of-action. METHODS AND RESULTS: Effects of WSSO on C. auris were tested by broth microdilution method, with observed IC50 at 5.96 mg ml-1. Time-kill assay revealed that WSSO is fungistatic. Mechanistically, ergosterol binding and sorbitol protection assays showed that C. auris cell membrane and cell wall are the targets for WSSO. Lactophenol: Cotton-Blue: Trypan-Blue staining confirmed loss of intracellular contents by WSSO treatment. Candida auris biofilm formation was disrupted by WSSO (BIC50: 8.52 mg ml-1). Additionally, WSSO exhibited dose and time-dependent mature biofilm eradication property with 50% efficacies at 23.27, 19.28, 18.18, and 7.22 mg ml-1 over 24, 48, 72, and 96 h, respectively. Biofilm eradication by WSSO was further substantiated through scanning electron microscopy. Standard-of-Care Amphotericin B, at its break-point concentration, (2 µg ml-1) was found to be inefficient as an antibiofilm agent. CONCLUSIONS: WSSO is a potent antifungal agent effective against planktonic C. auris and its biofilm.

Miconazole for the treatment of vulvovaginal candidiasis. In vitro, in vivo and clinical results. Review of the literature.

At concentrations achieved following systemic administration, the primary effect of imidazoles and triazoles on fungi is inhibition of 14-α-sterol demethylase, a microsomal cytochrome P450 (CYP) enzyme. Imidazoles and triazoles impair the biosynthesis of ergosterol for the cytoplasmic membrane and lead to the accumulation of 14-α-methyl sterols. The synthetic imidazole miconazole is additionally able to increase intracellular reactive oxygen species, at least in part through inhibition of fungal catalase and peroxidase. This unique feature of miconazole is probably the basis for its fungicidal activity in C. albicans, in addition to the fungistatic mode of action. Studies show that miconazole is superior to nystatin treatment and demonstrate its impact as one of the best options in managing vulvovaginal candidiasis. Regarding recurrent vulvovaginal candidiasis, several new drugs are currently developed to ensure effective treatment also for this group of patients.

Role of the nonribosomal peptide synthetase siderophore enzyme (Rfs) of Mucor lusitanicus in controlling the growth of fungal phytopathogens.

Dimorphic species of Mucor, which are cosmopolitan fungi belonging to subphylum Mucoromycotina, are metabolically versatile. Some species of Mucor are sources of biotechnological products, such as biodiesel from Mucor circinelloides and expression of heterologous proteins from Mucor lusitanicus. Furthermore, Mucor lusitanicus has been described as a model for understanding mucormycosis infections. However, little is known regarding the relationship between Mucor lusitanicus and other soil inhabitants. In this study, we investigated the potential use of Mucor lusitanicus as a biocontrol agent against fungal phytopathogens, namely Fusarium oxysporum f. sp. lycopersici, Fusarium solani, and Alternaria solani, which destroy economically important crops. Results showed that aerobic cell-free supernatants of the culture broth (SS) from Mucor lusitanicus inhibited the growth of the fungal phytopathogens in culture, soil, and tomato fruits. The SS obtained from a strain of Mucor lusitanicus carrying the deletion of rfs gene, which encodes an enzyme involved in the synthesis of siderophore rhizoferrin, had a decreased inhibitory effect against the growth of the phytopathogens. Contrarily, this inhibitory effect was more evident with the SS from an rfs-overexpressing strain compared to the wild-type. This study provides a framework for the potential biotechnological use of the molecules secreted from Mucor lusitanicus in the biocontrol of fungal phytopathogens.

Investigating the Resistance Mechanism of Wheat Varieties to Fusarium Head Blight Using Comparative Metabolomics.

Fusarium head blight (FHB) is primarily caused by Fusarium graminearum and severely reduces wheat yield, causing mycotoxin contamination in grains and derived products. F. graminearum-secreted chemical toxins stably accumulate in plant cells, disturbing host metabolic homeostasis. We determined the potential mechanisms underlying FHB resistance and susceptibility in wheat. Three representative wheat varieties (Sumai 3, Yangmai 158, and Annong 8455) were inoculated with F. graminearum and their metabolite changes were assessed and compared. In total, 365 differentiated metabolites were successfully identified. Amino acids and derivatives, carbohydrates, flavonoids, hydroxycinnamate derivatives, lipids, and nucleotides constituted the major changes in response to fungal infection. Changes in defense-associated metabolites, such as flavonoids and hydroxycinnamate derivatives, were dynamic and differed among the varieties. Nucleotide and amino acid metabolism and the tricarboxylic acid cycle were more active in the highly and moderately resistant varieties than in the highly susceptible variety. We demonstrated that two plant-derived metabolites, phenylalanine and malate, significantly suppressed F. graminearum growth. The genes encoding the biosynthetic enzymes for these two metabolites were upregulated in wheat spike during F. graminearum infection. Thus, our findings uncovered the metabolic basis of resistance and susceptibility of wheat to F. graminearum and provided insights into engineering metabolic pathways to enhance FHB resistance in wheat.

Fungistatic Activity Mediated by Volatile Organic Compounds Is Isolate-Dependent in Trichoderma sp. "atroviride B".

Trichoderma spp. produce multiple bioactive volatile organic compounds (VOCs). While the bioactivity of VOCs from different Trichoderma species is well documented, information on intraspecific variation is limited. The fungistatic activity of VOCs emitted by 59 Trichoderma sp. "atroviride B" isolates against the pathogen Rhizoctonia solani was investigated. Eight isolates representing the two extremes of bioactivity against R. solani were also assessed against Alternaria radicina, Fusarium oxysporum f. sp. lycopersici and Sclerotinia sclerotiorum. VOCs profiles of these eight isolates were analyzed using gas chromatography-mass spectrometry (GC-MS) to identify a correlation between specific VOCs and bioactivity, and 11 VOCs were evaluated for bioactivity against the pathogens. Bioactivity against R. solani varied among the fifty-nine isolates, with five being strongly antagonistic. All eight selected isolates inhibited the growth of all four pathogens, with bioactivity being lowest against F. oxysporum f. sp. lycopersici. In total, 32 VOCs were detected, with individual isolates producing between 19 and 28 VOCs. There was a significant direct correlation between VOC number/quantity and bioactivity against R. solani. 6-pentyl-α-pyrone was the most abundant VOC produced, but 15 other VOCs were also correlated with bioactivity. All 11 VOCs tested inhibited R. solani growth, some by >50%. Some of the VOCs also inhibited the growth of the other pathogens by >50%. This study demonstrates significant intraspecific differences in VOC profiles and fungistatic activity supporting the existence of biological diversity within Trichoderma isolates from the same species, a factor in many cases ignored during the development of biological control agents.

Birch Tar Introduced into Polylactide and Its Influence on the Barrier, Thermal, Functional and Biological Properties of the Film Obtained by Industrial Extrusion.

The aim of the study was to evaluate possibility of producing a polylactide film with birch tar by the industrial extrusion method and whether the addition of 10% birch tar can ensure adequate biocidal properties of PLA against pathogenic microorganisms (E. coli, S. aureus, P. aeruginosa, A. tumefaciens, X. campestris, P. brassicacearum, P. corrugate and P. syringae) and fungi (A. niger, A. flavus and A. versicolor) while ensuring beneficial functional properties, such as water vapor, nitrogen, oxygen and carbon dioxide permeability, which are of considerable importance in the packaging industry. The main test methods used were ISO 22196, ISO 846, ISO 2556, ASTM F 1927 and ASTM F 2476-20. The obtained results prove the possibility of extruding polymer films with a biocidal additive, i.e., birch tar, and obtaining favorable properties that qualify the produced film for applications in the packaging industry.

Analysis and control of fungal deterioration on the surface of pottery figurines unearthed from the tombs of the Western Han Dynasty.

In April 2020, 232 tombs of the Western Han Dynasty were found in Sundayuan, Heze City. In total, 141 pottery figurines of significant historical, cultural, and artistic value were unearthed from the tombs. Some of the figurines are currently being stored in warehouses, and the surface of some of the figurines show fungal deterioration. To thoroughly analyze the fungal deterioration on the surface of the pottery figurines and find appropriate control methods, we used high-through sequencing, scanning electron microscopy observation, pure cultures of culturable fungi, and optical microscopy observation and molecular identification of culturable fungi. We conducted fungistatic and simulation experiments in the laboratory to find appropriate control methods. We found that the fungi on the surface of the figurines were mainly of the phylum Ascomycota, and a few fungi were of the phyla Basidiomycota and Mucoromycota. We isolated seven culturable fungal strains and observed their colony morphology. The seven fungal strains were Lecanicillium aphanocladii, Penicillium aurantiogriseum, Clonostachys rosea, Mortierella sp., Mortierella alpina, Aspergillus flavus, and Cladosporium halotolerans. Through the fungistatic and simulation experiments conducted in the laboratory, we found that 50 mg/ml cinnamaldehyde and 0.5% K100 (2-methyl-4-isothiazolin-3-one) have a good fungistatic effect. They can not only inhibit the growth of fungi on medium, but also inhibit the growth of fungi on the surface of pottery figurines. This study has good reference significance for the analysis and control of fungal deterioration of unearthed pottery figurines.

Ag2O Nanoparticles as a Candidate for Antimicrobial Compounds of the New Generation.

Antibiotic resistance in microorganisms is an important problem of modern medicine which can be solved by searching for antimicrobial preparations of the new generation. Nanoparticles (NPs) of metals and their oxides are the most promising candidates for the role of such preparations. In the last few years, the number of studies devoted to the antimicrobial properties of silver oxide NPs have been actively growing. Although the total number of such studies is still not very high, it is quickly increasing. Advantages of silver oxide NPs are the relative easiness of production, low cost, high antibacterial and antifungal activities and low cytotoxicity to eukaryotic cells. This review intends to provide readers with the latest information about the antimicrobial properties of silver oxide NPs: sensitive organisms, mechanisms of action on microorganisms and further prospects for improving the antimicrobial properties.

Nanohybrids of oxides nanoparticles-chitosan and their antimicrobial properties.

Growing international problem with pathogens acquiring resistance to antibiotics is the reason for the search for bactericidal substances against which microorganisms cannot become resistant. The aim of this study was to synthesize inorganic-organic nanohybrids and obtain materials with antimicrobial effects. Chitosan (CS) was deposited on nanocomposite carriers such as calcium oxide with titanium dioxide (CaO-TiO2), magnesium oxide with titanium dioxide (MgO-TiO2) and copper(II) oxide with titanium dioxide (CuO-TiO2). The efficiency of the process was examined at varying concentrations of chitosan and temperature. The parameters for nanohybrids synthesis were selected based on the highest amount of nano-chitosan deposited on the nanohybrids-for each carrier, the process conditions were as follows: chitosan solution at 5 g l-1and 20 °C. The materials were obtained using these parameters and were used for microbiological tests againstE. coliATCC 25922,S. aureusATCC 25923 andC. albicansATCC 10231. The growth inhibitory activity of the obtained materials was qualitatively defined. These results suggest that the synthesized nanohybrids and nanocomposites exhibit biostatic action. The material with the broadest effect was the CuO-TiO2-CS hybrid, which had biostatic properties against all tested strains at a minimal concentration of 1250µg ml-1. Further research is required to find eco-friendly, non-toxic, and more effective antimicrobials with a broad action to prevent the acquisition of resistance.

Fungistatic Films Containing Cinnamon Essential Oil: New Coatings to Preserve the Nutraceutical Content of Avocado Fruit against Fusariosis.

Avocadoes are a rich source of nutrients and nutraceuticals that preserve human health. Nevertheless, this fruit is susceptible to phytopathogen infection during the postharvest period causing severe economic losses. Herein, we report on the in situ antifungal assessment of biodegradable films impregnated with the essential oil of Cinnamomum verum (CvEO) as natural fungistatic coatings to extend postharvest quality of Hass avocadoes (Persea americana cv. Hass). These coatings were evaluated on fruits previously infected with a native strain of Fusarium verticillioides. The cytotoxic assessment of CvEO on F. verticillioides revealed a minimum inhibitory concentration of 0.3±0.0 g L-1 whereas its chemical profiling showed (E)-cinnamaldehyde (45.9 %) 1,3,8-p-menthatriene (7.9 %) and linalool (6.8 %) as the major bioactive compounds. Four coatings (FC1-FC4) were made using chitosan (1 %) and a diverse quantity of CvEO (0.4-1.3 % w/v). The physicochemical properties demonstrated that the films FC3 and FC4 have the optimal characteristics of a food coating. Avocadoes preserved with the films FC3 and FC4 substantially kept (p<0.01) fruit firmness and the nutraceutical content of infected avocados at least for 21 d. The results of the present investigation suggest that our hybrid materials can conserve basic nutritional parameters such as fiber, protein, reducing sugars, as well as the content of unsaturated fatty acids which are the main nutraceuticals of this fruit.

Characterization of Fusarium acuminatum: A Potential Enniatins Producer in Tunisian Wheat.

Fusarium Head Blight (FHB), caused by multiple species of Fusarium in small grain cereals, is a significant and long-standing problem anywhere in the world. Knowing regional Fusarium spp. present on non-symptomatic grains and their potential for mycotoxin production is of concern for identifying novel actions for FHB and mycotoxin management, such as treatments with essential oils. Analyzing the mycotoxin content of grains from non-symptomatic ears of different wheat varieties cultivated in Tunisia, we isolated Fusaria specimens identified as F. culmorum and F. acuminatum using analysis of the partial DNA sequence of the ß-tubulin gene and ITS region. Two isolates of the latter species, uncommon in cereal grains in this region until now, were shown to be effective producers of enniatins in vitro, with 1390 and 3089 µg g-1 mycelial biomass (dry) in 11-day-old cultures. The susceptibility of an isolate of F. acuminatum to the fungistatic and antimycotoxin effects of eight essential oils was measured. Essential oils from Ammoides pusilla and Thymus capitatus used at 0.1 µL mL-1 in an agar culture medium, affected the mycelial growth by 55% and 79%, respectively and reduced the accumulation of enniatins per unit of mycelial colony by 26% and 52%, respectively. Finally, F. acuminatum was shown to be a contaminant of wheat grains in Tunisia and it may contribute to the contamination in enniatins. Two essential oils of Tunisian plants could be used for developing a biofungicide limiting both its mycelial growth and its accumulation of mycotoxins in grains.