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Penicillium expansum is the causal agent of blue mold in harvested fruits and vegetables during storage and distribution, causing serious economic loss. In this study we seek the action modes of bifonazole against this pathogen. Bifonazole exhibited strong antifungal activity against P. expansum by inhibiting ergosterol synthesis. The ergosterol depletion caused damage to the cell structure and especially cell membrane integrity as observed by SEM and TEM. With increased unsaturated fatty acids contents, the cell membrane viscosity decreases and can no longer effectively maintain the cytoplasm, which ultimately decreases extracellular conductivity, changes intracellular pH and ion homeostasis. Exposure of hyphal cells to bifonazole shows that mitochondrial respiration is inhibited and reactive oxygen species (ROS) levels–including H2O2 and malondialdehyde (MDA) – are significantly increased. The functional impairment of mitochondria and cell membrane eventually cause cell death through intrinsic apoptosis and necroptosis.

It is not clear how the amino acid composition of the X sites in GXSXG domain affects the properties of the lipase. To address this, saturation mutation at these sites of Penicillium expansum lipase (PEL) was carried out. After screening, a mutant L133M with significantly different activity was obtained. The activity of L133M was lower than that of wild-type PEL (WT) when the reaction time was 3–10 min. However, it was 1.61 times that of WT under long-term (48 h) catalysis. WT exhibited interfacial activation when the concentration of p-nitrophenylbutyrate was higher than 3 mM, whereas L133M could only show interfacial activation at higher micelle concentration (p-nitrophenylbutyrate concentration > 5 mM), which seemed to be slower. Further analysis showed that a new hydrophobic interaction was generated between the methionine introduced at position 133 and the lid, which made the interface activation process more difficult. However, the binding energy between the activated mutant and the substrate was smaller, showing stronger activity. This work provides some reliable clues for further understanding the complex role of GXSXG domain in enzyme catalysis.

The rise in world industrialization and the cost of importing enzyme by local industries have led to arise in the search for novel and native enzyme producing microorganisms. Cellulase is an enzyme that catalyzes the breaking down of carbon chains in cellulose and hemicellulose, this research therefore aimed at studying fungal cellulase produced by Penicillium expansum grown on malus domestica (apple fruits). Fresh apple fruit was allowed to deteriorate under laboratory condition until there was visible mould growth. The mould with desired features of the organism of interest was subcultured by direct plating on PDA plates to which 10 % streptomycin has been added to prevent bacterial contaminants. The plates were incubated at 28±2 0C for 7 days until a visible mass of blue mycelia appear. The isolate was further subcultured onto freshly prepared media until pure culture was obtained. Characterization and identification of isolate were done using macroscopy and microscopy techniques. The isolate was re-inoculated into healthy apple fruits and the fruits were incubated at temperature of 28±2 oC for 8 days. Cellulolytic activity was examined every day throughout the incubation period. Crude enzyme was extracted each day using standard methods. Carboxyl methyl cellulose was used as standard for the crude cellulase activity assay after extraction from the infected apple fruits using Dinitrosalicylic acid (DNSA). Culture parameters like pH and temperature were also optimized to determine their effect on cellulolytic activity of the fungus. Cellulase activity was defined as the amount of glucose produced in μmol/mg/min under the assay condition. The highest cellulase activity of 86.84±0.52 μmol/mg/min was observed on day 6 of incubation at 28±2 oC and at pH 7. In conclusion, it is evident from this research that P. expansum isolated could be used as potential novel organism for industrial production of cellulase under optimized fermentation conditions.

The mycotoxin patulin is primarily produced as a secondary metabolite by numerous fungal species and predominantly by Aspergillus, Byssochlamys, and Penicillium species. It is generally associated with fungal infected food materials. Penicillium expansum is considered the only fungal species liable for patulin contamination in pome fruits, especially in apples and apple-based products. This toxin in food poses serious health concerns and economic threat, which has aroused the need to adopt effective detection and mitigation strategies. Understanding its origin sources and biosynthetic mechanism stands essential for efficiently designing a management strategy against this fungal contamination. This review aims to present an updated outline of the sources of patulin occurrence in different foods and their biosynthetic mechanisms. It further provides information regarding the detrimental effects of patulin on human and agriculture as well as its effective detection, management, and control strategies.

Disease incidence and disease severity of avocado fruit rot were assessed on cv. Hass and cv. Fuerte during 2019 and 2020 growing seasons. The highest percentages of disease incidence and disease severity were recorded in cv. fuerte in the market at El-Qanater El-Khayria, Qaliobia Governorate. On the other hand, the lowest percentages of disease incidence and disease severity were recorded in cv. Hass in the field at El- Rahawy, Giza Governorate during the two growing seasons. Four different fungi, i.e., Colletotrichum gloeosporioides, Lasiodiplodia theobromae, Fusarium moniliforme and Penicillium expansum were isolated from the naturally infected avocado fruits in the surveyed regions. Pathogenicity tests showed that the maximum percentage of disease severity on cv. Hass and cv. Fuerte resulted from infection by L. theobromae, followed by C. gloeosporioides, while the lowest percentage of disease severity was incited byP. expansum. The efficacy of alternative products namely essential oils of Lemongrass (Cymbopogon citratus L.), Rosemary (Rosmarinus officinalis L.) and Thyme (Thymus vulgaris L.) and Serenade ASO (Bacillus subtilis QST713) was evaluated in in vitro and in vivo as well as under field conditions for controlling fruit rot of avocado comparing with Imazalil, Copper oxide and Copper oxide nanoparticles. Complete inhibition of colony growth and protection from fruit rot was recorded with essential oils and Copper oxide at concentration of 3 ml or g/L of each. Meanwhile, the same effect was recorded at concentration of 1 ml/ L in case of Imazalil, Copper oxide nanoparticles and Serenade ASO. However, under field conditions, Copper oxide nanoparticles and Imazalil gave the highest efficacy for controlling avocado fruit rot disease, followed by Serenade ASO, Copper oxide and Thyme, while the lowest efficacy was recorded when trees sprayed with Lemongrass and Rosemary, respectively. The results indicated that avocado fruit rot disease could be controlled using suitable concentration of the tested treatments.

BACKGROUND: Postharvest diseases in fruits and vegetables are one of the major problems in storing them as a fresh agri-product. This study aimed to investigate the antifungal activity of licorice (Glycyrrhiza glabra) aqueous extract against the Penicillium expansum and the Penicillium digitatum in apple and tangerine fruits as well as their postharvest decay during storage time. METHODS: The minimum inhibitory concentration (MIC) of the molds, and the decay inhibition percentage (%DI) with the P.expansum for apple and P.digitatum for tangerine after treatment with licorice aqueous extract were measured. Additionally, the lesion diameter, titratable acidity (TA), total soluble solids (TSS), pH, and organoleptic properties were determined. RESULTS: The growth of molds was almost inhibited at the concentration of 62.5 mg/mL. The ability of licorice aqueous extract to significantly control and reduce the growth of P. expansum in apple by 60 and 20 % after 7 days and 21 days of storage time was proved, respectively. Furthermore, significant differences in pH and TSS (p

Mold growth is among the major causes of health impairment of cereals, in particular durum wheat (Triticum durum) for the synthesis of mycotoxins such as aflatoxins B1 and ochratoxin A (OTA), originally from poisoning in the consumer. In this context, the objectives of this work is the search and characterization of fungal strains Producers mycotoxins such as Aspergillus, Penicillium in semolina and their derivatives (traditional and industrial couscous) and detect and quantify total aflatoxins, aflatoxins B1 and ochratoxin mycotoxicologique to assess the risk associated with the consumption of these foods. In this regard, our work focuses on mycological and mycotoxicologique study of semolina and couscous deemed most commercialized in the town of Bechar-Algeria after a socio-economic survey. The mycological study testifies the high degree of pollution of our samples by Aspergillus, Penicillium. The expertise of genera reveals the high degree of invasion of our samples by Aspergillus, Penicillium. The examination of fungal procession characterizing our samples shows a very high index of distribution, or of fidelity of Penicillium 43.75% of our sample and 28.38% Aspergillus. The presence of these species is evidence that our samples have been abused, but especially poorly stored; should be noted the involvement of the genera Alternaria 7.10%, Fusarium 13.70%. Thin-layer chromatographic (TLC) analysis revealed that 50% of Aspergillus flavus-parasiticus strains were aflatoxin G-producing and aflatoxin B-free in our samples. Of the Aspergillus ochraceus strains 50% were OTA producers. The presumption of toxicity of the various samples appeared positive on TLC. The test of Elisa has confirmed the presence of the OTA in our samples, the analysis of its results shows that the majority of the rates of OTA taken on our analyzed samples follow the European standard, these rates are between 1.01 and 1.9, except for one sample (couscous) which has shown a rate much higher than the standard recommended by the regulation (> 100 ppb), the samples of semolina had a rate of OTA lower than the beginning of the detection (1 ppb). The results of the presence of AFB spread out between 4.93 ppb and > 40 ppb. The antifungal activity of the resin of Boswellia carterii was tested on the following strains: Aspergillus niger, Aspergillus flavus, Penicillium expansum. And kneaded according to the technique of diagonal growth on intermediate solid medium (PDA). The results showed that the yield of the aqueous extract varied between 96.2 and 99.8%. The results of the extracts also showed activity against the fungi studied 48.6% and 96.2%.

Penicillium expansum Link causes an economically important postharvest blue mold disease in a number of fruits and vegetables. In the present study, this fungus was isolated from rotted grapes and identified on morphological basis. Identification of the pathogen was further confirmed on molecular basis by using four different primer pairs namely ITS, β-tubulin, CMD and CF under accession numbers MN752155, MN787831, MN787832 and MN787833, respectively. Leaf extract of Chenopodium murale was assessed for its potential to control in vitro growth of P. expansum. For this purpose, leaves were extracted in methanol and after evaporation of the solvent, the resulting extract was successively partitioned with n- hexane, chloroform, ethyl acetate and n- butanol followed by antifungal bioassays with different concentrations (1.562 to 200 mg mL -1 ) each organic solvent fraction. Although all the fractions variably controlled the fungal pathogen, however, n- butanol fraction showed the highest antifungal activity causing 45–86% reduction in biomass of the pathogen. Ethyl acetate fraction was also highly antifungal and reduced fungal biomass by 44–81%. Chloroform and n- hexane fractions were comparatively less effective and reduced biomass of P. expansum by 30–72% and 11–44%, respectively. This study concludes that ethyl acetate and n- butanol fractions are highly antifungal in nature against P. expansum.

Penicillium is one of the most important postharvest pathogens of citrus fruits worldwide. It induces blue or green mold disease, a decay that can lead to significant economic losses during storage. Based on internal transcribed spacer (ITS) sequences, seven Penicillium species and one closely related Talaromyces variabilis were identified from 30 rotten samples of citrus fruits marketed in Qena. Penicillium expansum was the most common species, recovered from 16.7% of the samples, followed by P. chrysogenum (10%) and P. polonicum (10%). Sixteen isolates were tested through inoculation on healthy citrus fruits; the data exhibited that 68.7% of isolates were highly virulent. A “Specific Gene Random Primer Polymerase Chain Reaction (SGRP-PCR)” marker technique indicated that the genetic similarity among P. expasum ranged from 49.4 to 85.7%, and a relatively correlation was found between SGRP band profile and species origin. Patulin was detected in 40% of P. expansum isolates. This study provided a useful molecular approach to identify different Penicillium species by sequencing ITS region, focus on the pathogenicity, compare between P. expansum isolates and their ability in patulin production.

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.

In this study, green and eco-friendly biosynthesis of selenium nanoparticles (Se-NPs) were performed using Penicillium expansum ATTC 36200 for multiple biomedical applications. Mycosynthesized Se-NPs were completely characterized using UV, FT-IR, XRD, SEM, and TEM techniques. Se-NPs biosynthesized by P. expansum was characterized as a spherical shape with average size 4 to 12.7 nm. Moreover, Se-NPs were evaluated for multiple biomedical applications as antimicrobial, antioxidant, and anticancer activities and hemocompatibility. Results illustrated that Se-NPs have potential antimicrobial activity against Gram-positive (Bacillus subtilis ATCC6051 and Staphylococcus aureus ATCC23235), Gram-negative bacteria (Escherichia coli ATCC8739and Pseudomonas aeruginosa ATCC9027), fungi (Candida albicans ATCC90028, Aspergillus niger RCMB 02724 and Aspergillus fumigatus RCMB 02568), and antioxidant activity. Additionally, Se-NPs exhibited anticancer activity against PC3 cell line; IC50 was 99.25 μg/mL. Meanwhile, they showed non-hemolytic activity on human RBCs at concentration up to 250 μg/mL. In conclusion, biosynthetic Se-NPs by P. expansum are promising for many safe-use biomedical applications.

Among the postharvest pathogens of apple fruit, Penicillium expansum (P.expansum) is considered to be one of the most common fungal pathogens worldwide. It is the causal agent of blue mold decay; the main serious economically disease of apple fruits, and the main producer of patulin (PAT). The principal objective of this work was to evaluate the aggressiveness and the PAT production of three isolated strains from apples produced in the East of Algeria. These strains were isolated then screened for their inhibitory activity against Escherichia coli and Bacillus subtilis (18 mm and 18 mm for S7; 10 mm and 11mm for S3), as well as their ability to cause blue rot (S4). The three strains S7, S3, and S4 (MN904448, MT023795 and MN904449, respectively) were identified as P. expansum by ITS1, ITS4, and β-tubulin gene sequences analysis. The in vitro ability to produce PAT was confirmed by Gas chromatography-mass spectrometry of strains’ crude extracts cul-tured on YES (Yeast-Extract-Sucrose) medium. Indeed, these strains exhibited a great capacity to cause blue mold decay for all tested apples (26 to 52 mm lesion diameter), after nine days of incubation at 25°C. Apples rotten area analysis that was performed by UPLC analysis revealed that these three strains were able to produce PAT at different concentrations ranging from 0.57 to 469 μg/mL. Thus, these strains constitute a risk to consumer health because these amounts exceed the permissive level of PAT.

A total of 13 samples of grapes (bunches) without apparent fungal contamination were analyzed. The samples were collected during the 2019 harvest from Vrbové village in the Small Carpathian region of Slovakia. For the isolation of fungi were used the direct plating technique on DRBC plates. The plates were incubated aerobically at 25 ±1 °C for one week in the dark. The data obtained from the cultivation of the grape berry samples revealed a high diversity of fungal species (a total of 1044 isolates were obtained). Alternaria and Rhizopus were the main components of the wine grape mycobiota of the Vrbovský subregion at harvest time (92%, each), followed by Cladosporium (85%), Penicillium (77%), Botrytis and Epicoccum (54%, each). The most abundant genera found by descending order were Penicillium (25%), Alternaria (24%), Cladosporium (20%), and Rhizopus (12%) and only in minor percentage by Aspergillus (3%) among others. The main fungal species isolated from genera Penicillium and Aspergillus were Penicillium expansum (57% RD) and A. section Nigri (97% RD). Of 17 analyzed Penicillium strains, 65% were able to produce at least one of the six mycotoxins analyzed in in vitro conditions by means of thin-layer chromatography method: citrinin, griseofulvin, patulin, cyclopiazonic acid, penitrem A, and roquefortin C.

Blue mould, caused primarily by Penicillium expansum, is a major threat to the global pome fruit industry, causing multimillion‐dollar losses annually. The blue mould fungus negatively affects fruit quality, thereby reducing fresh fruit consumption, and significantly contributes to food loss. P. expansum also produces an array of mycotoxins that are detrimental to human health. Management options are limited and the emergence of fungicide‐resistant Penicillium spp. makes disease management difficult, therefore new approaches and tools are needed to combat blue mould in storage. This species profile comprises a comprehensive literature review of this aggressive pathogen associated with pomes (apple, pear, quince), focusing on biology, mechanisms of disease, control, genomics, and the newest developments in disease management. Taxonomy Penicillium expansum Link 1809. Domain Eukaryota, Kingdom Fungi, Phylum Ascomycota, Subphylum Pezizomycotina, Class Eurotiomycetes, Subclass: Eurotiomycetidae, Order Eurotiales; Family Trichocomaceae, Genus Penicillium, Species expansum. Biology A wide host range necrotrophic postharvest pathogen that requires a wound (e.g., stem pull, punctures, bruises, shoulder cracks) or natural openings (e.g., lenticel, stem end, calyx sinus) to gain ingress and infect. Toxins Patulin, citrinin, chaetoglobosins, communesins, roquefortine C, expansolides A and B, ochratoxin A, penitrem A, rubratoxin B, and penicillic acid. Host range Primarily apples, European pear, Asian pear, medlar, and quince. Blue mould has also been reported on stone fruits (cherry, plum, peach), small fruits (grape, strawberry, kiwi), and hazel nut. Disease symptoms Blue mould initially appears as light tan to dark brown circular lesions with a defined margin between the decayed and healthy tissues. The decayed tissue is soft and watery, and blue‐green spore masses appear on the decayed area, starting at the infection site and radiating outward as the decayed area ages. Disease control Preharvest fungicides with postharvest activity and postharvest fungicides are primarily used to control decay. Orchard and packinghouse sanitation methods are also critical components of an integrated pest management strategy. Useful websites Penn State Tree Fruit Production Guide (https://extension.psu.edu/forage‐and‐food‐crops/fruit), Washington State Comprehensive Tree Fruit (http://treefruit.wsu.edu/crop‐protection/disease‐management/blue‐mold/), The Apple Rot Doctor (https://waynejurick.wixsite.com/applerotdr), penicillium expansum genome sequences and resources (https://www.ncbi.nlm.nih.gov/genome/browse/#!/eukaryotes/11336/)., This article is a synthesis and compilation of the latest information on the mycotoxingenic blue mould fungus from multiple perspectives that entail omics, biology, and tools for decay control.

Penicillium expansum is a necrotrophic plant pathogen with a wide range of fruit hosts. It causes blue mold rot during fruit storage, transport, and sale, resulting in huge economic losses to the fruit industry. Moreover, this pathogen is also the main producer of patulin, a toxic secondary metabolite that contaminates fruit and fruit-derived products and impairs human health. Therefore, understanding molecular basis of the pathogenicity and patulin biosynthesis of the fungal pathogen has important scientific significance and also plays an important guiding role in the research and development of new control technologies. Here, we comprehensively summarize the recent research progress, particularly regarding the molecular aspects of pathogenicity, patulin biosynthesis, and the related regulatory mechanisms, as well as control technologies for blue mold rot in the fruit industry.

Nowadays, there is an increasing concern regarding the shelf life of food products, leading producers to research natural antimicrobial agents to use in food preparation. In this study, we evaluated the antifungal activity of Lactobacillus plantarum fermented whey and then added the whey during preparation of pita bread to study shelf-life improvement. The fermented whey showed a satisfactory inhibitory (antifungal) effect against Penicillium expansum and Penicillium brevicompactum strains: the minimum inhibitory and minimum fungicidal concentrations ranged from 3.9 to 39.0 g/L and from 62.5 to 250 g/L, respectively. Addition of fermented whey increased the shelf life of the pita bread. After inoculation of the bread surface with Penicillium, an increase in shelf life until d 8 was achieved compared with the positive control, whereas under natural contamination conditions, an extension of shelf life until d 19 was observed. In terms of antimicrobial activity, the greatest reduction (100%) in fungal growth was achieved when all of the water in the dough was replaced with fermented whey. An untrained sensory panel could not identify differences between bread produced with fermented whey and control pita breads. These results suggest the possibility of using fermented whey in food preservation.

Aims To conduct biological investigations and to evaluate the antimicrobial and antioxidant activities of the essential oils (EOs) extracted from Juniperus communis, J. scopulorum and J. horizontalis; to screen their mechanisms of action by conducting the cell membrane permeability assay (CMP); and to determine the possible cytotoxicity of the three EOs against human neuroblastoma cells (SH-SY5Y). Methods and results The antifungal activity was tested against four phytopathogenic fungi (Monilinia fructicola, Aspergillus niger, Penicillium expansum and Botrytis cinerea). The antibacterial activity was evaluated against two Gram-positive (G+ve) (Bacillus megaterium and Clavibacter michiganensis) and three Gram-negative (G-ve) bacterial strains (Pseudomonas fluorescens, P. syringae pv. phaseolicola and Xanthomonas campestris). Results showed that the three tested EOs have antifungal activity against M. fructicola and P. expansum and effective antibacterial activity against P. syringae pv. phaseolicola and B. megaterium. Moreover, the three EOs were evaluated for their ability to inhibit the growth of SH-SY5Y cells with MTT assay. J. communis EO was the more effective with an IC50 of 53·7 μg ml-1 . The antioxidant capacity of the three EO did not differ as measured by the DPPH assay. Conclusions The three tested juniper EOs showed promising antimicrobial and antioxidant activity and cytotoxic effects against human neuroblastoma cell line. Significance and impact of the study The outfindings from this research showed promising antimicrobial effects of the three oils against the majority of the tested phytopathogens with a potential to utilize them as natural alternatives to synthetic drugs, the cause of global environmental problems, pathogen resistance and difficulty to control many post-harvest plant diseases.

Antifungal compounds are of interest to reduce commodity spoilage and exposure to mycotoxins. In this study, a series of quantitative structure-activity relationship (QSAR) equations based on topological properties were developed to gain insight into the antifungal activities of phenolic compounds. The molecules were geometry optimized using B3LYP/6-311+G** density functional theory calculations. Analysis of the frontier orbital properties revealed that conjugated phenolic compounds possessed smaller band gap energies. Genetic function approximation (GFA) on populations of 100 one to two descriptor models over 10,000 generations identified several models for antifungal activity against Fusarium verticillioides, Fusarium oxysporum, Aspergillus flavus, Aspergillus fumigatus, Penicillium expansum, and Penicillium brevicompactum. Phenolic compounds with greater antifungal activity possessed a lower electrophilicity index. The correlation coefficients for the one and two descriptor models ranged from 0.627 to 0.790 and 0.762 to 0.939, respectively. Molecular descriptors associated with electrostatic and topological properties are important to describe the antifungal activities of the phenolic compounds studied.

Bu çalışmada, borik asitin Penicillium expansum’a karşı etkinliği hem in vitro hem de in vivo koşullarda değerlendirilmiştir. In vitro’da, borik asitin %0.125 konsantrasyonu P. expansum’un misel gelişmesini, spor çimlenmesini ve çim tüpü uzunluğunu sırasıyla %68.88, %74.00 ve %85.50 oranında azaltmış, halbuki daha yüksek konsantrasyonlar (%0.25, %0.50, %1.00 ve %2.00)’ı ise tamamen engellemiştir. Ayrıca borik asitin yukarıda belirtilen ilk konsantrasyonu ve sonraki konsantrasyonlarının etkileri arasındaki fark istatistiksel olarak önemli bulunmuştur (P%2 olarak belirlenmiştir. In vivo denemelerde, borik asitin %0.25 konsantrasyonu hariç, diğer tüm konsantrasyonlar (%0.50, %1.00 ve %2.00)’ı hem koruyucu hem de tedavi edici etki göstermişler ve bu konsantrasyonlar mavi küfün hastalık şiddetini patojen ile inokule edilmiş kontrole kıyasla önemli ölçüde azaltmıştır (P

Cassava peels are waste generated from cassava processing and are mostly disposed of in Nigeria by dumping indiscriminately in landfills/waste dumps where they remain as huge mounds and constitute nuisance to the environment. This study was carried out using cassava peels as substrate for the production of α – amylase from three fungi (Aspergillus flavus, Aspergillus niger and Penicillium expansum) selected from twelve microbial isolates obtained from biodegrading cassava peels. The fungi were isolated using cassava peel agar medium and the α – amylase was produced by the solid state fermentation process. The α – amylase was assayed by measuring the decrease in staining power of starch with iodine reagent at 620nm and purified by using Sephadex G-100 and Sephadex C-50. The α – amylase was characterized by examining the effect of temperature, stability at 70oC, pH, substrate concentration, metal ions and EDTA. The results show that the purification fold and specific activity were 95.727, 1.053 Units/mg protein; 81.830, 0.982 Units/mg protein and 85.784, 0.686 Units/mg protein for A. flavus, A. niger and P. expansum respectively. The optimum temperature and pH were 45oC and 4.5 respectively. It was observed that the α – amylase still retained some activity after heating at 70oC for 35 min. The α – amylase activity increased with increase in substrate concentration and metal ion concentration (Na+, K+, Mg2+ and Ca2+) but decreased with increase in heavy metal ion concentration (Hg2+ and Pb2+) and EDTA. Key words: Cassava peels, α – amylase, fungi, starch and Sephadex C – 50.

Antagonistic yeasts can inhibit fungal growth. In our previous research, Meyerozyma guilliermondii, one of the antagonistic yeasts, exhibited antagonistic activity against Penicillium expansum. However, the mechanisms, especially the molecular mechanisms of inhibiting activity of M. guilliermondii, are not clear. In this study, the protein expression profile and transcriptome characterization of P. expansum induced by M. guilliermondii were investigated. In P. expansum induced by M. guilliermondii, 66 proteins were identified as differentially expressed, among them six proteins were upregulated and 60 proteins were downregulated, which were associated with oxidative phosphorylation, ATP synthesis, basal metabolism, and response regulation. Simultaneously, a transcriptomic approach based on RNA-Seq was applied to annotate the genome of P. expansum and then studied the changes of gene expression in P. expansum treated with M. guilliermondii. The results showed that differentially expressed genes such as HEAT, Phosphoesterase, Polyketide synthase, ATPase, and Ras-association were significantly downregulated, in contrast to Cytochromes P450, Phosphatidate cytidylyltransferase, and Glutathione S-transferase, which were significantly upregulated. Interestingly, the downregulated differentially expressed proteins and genes have a corresponding relationship; these results revealed that these proteins and genes were important in the growth of P. expansum treated with M. guilliermondii.

The blue mold fungus, Penicillium expansum, is a postharvest apple pathogen that contributes to food waste by rotting fruit and by producing harmful mycotoxins (e.g. patulin). To identify genes controlling pathogen virulence, a random T-DNA insertional library was created from wild-type P. expansum strain R19. One transformant, T625, had reduced virulence in apples, blistered mycelial hyphae, and a T-DNA insertion that abolished transcription of the single copy locus in which it was inserted. The gene, Blistering1, encodes a protein with a DnaJ domain, but otherwise has little homology outside the Aspergillaceae, a family of fungi known for producing antibiotics, mycotoxins, and cheese. Because protein secretion is critical for these processes and for host infection, mass spectrometry was used to monitor proteins secreted into liquid media during fungal growth. T625 failed to secrete a set of enzymes that degrade plant cell walls, along with ones that synthesize the three final biosynthetic steps of patulin. Consequently, the culture broth of T625 had significantly reduced capacity to degrade apple tissue and contained 30 times less patulin. Quantitative mass spectrometry of 3,282 mycelial proteins revealed that T625 had altered cellular networks controlling protein processing in the endoplasmic reticulum, protein export, vesicle-mediated transport, and endocytosis. T625 also had reduced proteins controlling mRNA surveillance and RNA processing. Transmission electron microscopy of hyphal cross sections confirmed that T625 formed abnormally enlarged endosomes or vacuoles. These data reveal that Blistering1 affects internal and external protein processing involving vesicle-mediated transport in a family of fungi with medical, commercial, and agricultural importance.

Exogenous decanal can significantly inhibit the germination and development of postharvest pathogen Penicillium expansum in vitro. Through an iTRAQ-based analysis, a global view of proteomic alteration of P. expansum spores under decanal treatment was acquired. A total of 246 up-regulated and 293 down-regulated differentially expressed proteins (DEPs) were identified. Among them, DEPs related to glutathione metabolism, ribosome, and oxidative phosphorylation pathway were noticed for their functional significance, large number and high rich value in pathway enrichment statistics. Further analysis found that, under decanal stress, expression of 9 genes corresponding to DEPs involved in the oxidative phosphorylation pathway showed a significantly decreasing trend, and activities of 3 crucial enzymes (NADH dehydrogenase, CoQ-Cytochrome c reductase, and F1F0-ATP synthetase) were inhibited. Also, a decreased in ATP content, reduction in the number of mitochondria, and weakening in carbohydrate consumption were detected. Based on these results, disturbance of oxidative phosphorylation would partly be responsible for the inhibitory effect of decanal on the growth of P. expansum. The findings would provide new insights into exploring the possible antifungal mechanisms of decanal.

Red yeasts grow on food wastes, show antagonistic activity against food-spoilage microorganisms, produce food supplements, and may be utilized as feed-supplements themselves to positively modulate the quali-quantitative composition of intestinal microbiota. Therefore, they show a variety of possible biotechnological applications in agri-food-related processes. Here, to further explore the biotechnological potential of red yeasts, eleven strains ascribed to different species of the genera Rhodotorula and Sporobolomyces, differing for biomass and carotenoids production, were characterized in vitro for biofilm formation, invasive growth, and growth at the temperature range of 20–40 °C and in vivo for their antagonistic activity against the fungal pathogen and patulin producer Penicillium expansum. Most of them formed cellular MAT and showed invasive growth as well as adhesion to plastic materials. Four strains determined a significant reduction of fruit decay caused by P. expansum on apple fruit while the remaining seven showed different degrees of biocontrol activity. Finally, none of them grew at body temperature (>37 °C). Statistical analyses of both qualitative and quantitative phenotypic data, including biomass and carotenoids production, gathered further information on the most interesting strains for the biotechnological exploitation of red yeasts in agri-food-related process.

To the best of our knowledge, this is the first study demonstrating the efficiency of Allium sativum hydro-alcoholic extract (ASE) againstFigure growth, biofilm development, and soluble factor production of more than 200 biodeteriogenic microbial strains isolated from cultural heritage objects and buildings. The plant extract composition and antioxidant activities were determined spectrophotometrically and by HPLC–MS. The bioevaluation consisted of the qualitative (adapted diffusion method) and the quantitative evaluation of the inhibitory effect on planktonic growth (microdilution method), biofilm formation (violet crystal microtiter method), and production of microbial enzymes and organic acids. The garlic extract efficiency was correlated with microbial strain taxonomy and isolation source (the fungal strains isolated from paintings and paper and bacteria from wood, paper, and textiles were the most susceptible). The garlic extract contained thiosulfinate (307.66 ± 0.043 µM/g), flavonoids (64.33 ± 7.69 µg QE/g), and polyphenols (0.95 ± 0.011 mg GAE/g) as major compounds and demonstrated the highest efficiency against the Aspergillus versicolor (MIC 3.12–6.25 mg/mL), A. ochraceus (MIC: 3.12 mg/mL), Penicillium expansum (MIC 6.25–12.5 mg/mL), and A. niger (MIC 3.12–50 mg/mL) strains. The extract inhibited the adherence capacity (IIBG% 95.08–44.62%) and the production of cellulase, organic acids, and esterase. This eco-friendly solution shows promising potential for the conservation and safeguarding of tangible cultural heritage, successfully combating the biodeteriogenic microorganisms without undesirable side effects for the natural ecosystems.

Postharvest biocontrol agents are considered a viable alternative to the use of synthetic chemicals as demonstrated by extensive research conducted by scientists and companies worldwide. In the present investigation, the biocontrol potential of a carotenoid-producing basidiomycetous yeast isolated from table grape flowers was analyzed. The strain RY1 proved to be Sporobolomyces roseus. In vitro and in vivo tests were conducted to assess its efficacy against Penicillium expansum, one of the most important postharvest pathogens and producer of the mycotoxin patulin. The yeast proved to control both fungal growth and patulin production, and, in addition, to greatly affect disease incidence and severity on apples. Its mode of action is presumably related both to the competition for nutrients and the production of antifungal volatiles. As such, although further large-scale trials are needed, our S. roseus strain represents a potential interesting biocontrol agent to be applied after harvest.

Nowadays, increase fruit losses are being reported due to the development of fungal postharvest diseases. In an attempt to reduce the use of synthetic fungicides, a turn towards natural products such as essential oils (EOs) and natural compounds has been made. The objective of this study was to investigate the effects of eucalyptus (Euc), rosemary (Ros) EO, their mixture (50:50 v/v) and their common main component (i.e., eucalyptol) on the quality parameters, fruit response and inhibition of blue rot (Penicillium expansum) in apple and pear fruits during their shelf life. The results of the present study revealed that fungal colony growth decreased in vitro with exposure at eucalyptus EO (Euc-300 μL/L), rosemary EO (Ros-300 μL/L) and their mixture (Euc + Ros 100 and 300 μL/L). The exposure at Ros-100 μL/L stimulated spore production, whilst Euc + Ros (100 and 300 μL/L) and eucalyptol (100 and 300 μL/L) decreased spore germination. Moreover, the in vivo applied treatments resulted in decreased lesion growth of P. expansum in apple and pear fruits. Respiration rate increased with the application of Euc + Ros at 300 μL/L and eucalyptus EO (Euc-100 μL/L and Euc-300 μL/L) for both assessed fruits. On the other hand, no significant differences were reported on apples and pears total soluble solids and acidity values. The application of Euc + Ros-300 μL/L in apples increased hydrogen peroxide (H2O2) levels, whilst Euc-100 and Euc-300 μL/L increased lipid peroxidation levels. Regarding pear fruits, exposure to Euc-100 μL/L and Ros-100 μL/L resulted in increased H2O2 whereas, Euc-100 μL/L, Ros- (100 and 300 μL/L) and eucalyptol (100 and 300 μL/L) also increased lipid peroxidation. The findings of this study indicate that the investigated natural products can be explored for the preservation of fresh apples and pears, as alternative natural fungicides with consideration of the fresh produce quality attributes.

Penicillium expansum is a necrotrophic wound fungal pathogen that secrets virulence factors to kill host cells including cell wall degrading enzymes (CWDEs), proteases, and mycotoxins such as patulin. During the interaction between P. expansum and its fruit host, these virulence factors are strictly modulated by intrinsic regulators and extrinsic environmental factors. In recent years, there has been a rapid increase in research on the molecular mechanisms of pathogenicity in P. expansum, however, less is known regarding the bacteria–fungal communication in the fruit environment that may affect pathogenicity. Many bacterial species use quorum-sensing (QS), a population density-dependent regulatory mechanism, to modulate the secretion of quorum-sensing signaling molecules (QSMs) as a method to control pathogenicity. N-acyl homoserine lactones (AHLs) are Gram-negative QSMs. Therefore, QS is considered an antivirulence target, and enzymes degrading these QSMs, named quorum-quenching enzymes, have potential antimicrobial properties. Here, we demonstrate that a bacterial AHL lactonase can also efficiently degrade a fungal mycotoxin. The mycotoxin is a lactone, patulin secreted by fungi such as P. expansum. The bacterial lactonase hydrolyzed patulin at high catalytic efficiency, with a kcat value of 0.724 ± 0.077 s−1 and KM value of 116 ± 33.98 μM. The calculated specific activity (kcat/KM) showed a value of 6.21 × 103 s−1M−1. While the incubation of P. expansum spores with the purified lactonase did not inhibit spore germination, it inhibited colonization by the pathogen in apples. Furthermore, adding the purified enzyme to P. expansum culture before infecting apples resulted in reduced expression of genes involved in patulin biosynthesis and fungal cell wall biosynthesis. Some AHL-secreting bacteria also express AHL lactonase. Here, phylogenetic and structural analysis was used to identify putative lactonase in P. expansum. Furthermore, following recombinant expression and purification of the newly identified fungal enzyme, its activity with patulin was verified. These results indicate a possible role for patulin and lactonases in inter-kingdom communication between fungi and bacteria involved in fungal colonization and antagonism and suggest that QQ lactonases can be used as potential antifungal post-harvest treatment.

The apple is one of the most important fruit tree crops in the Mediterranean region. Lebanon, in particular, is among the top apple producer countries in the Middle East, however, recently, several types of damage, particularly rot symptoms, have been detected on fruits in cold storage. This study aims to identify the causal agents of apple decay in Lebanese post-harvest facilities and characterize a set of 39 representative strains of the toxigenic fungus Penicillium. The results demonstrated that blue mould was the most frequent fungal disease associated with apples showing symptoms of decay after 3–4 months of storage at 0 °C, with an average frequency of 76.5% and 80.6% on cv. Red and cv. Golden Delicious apples, respectively. The morphological identification and phylogenetic analysis of benA gene showed that most Penicillium strains (87.2%) belong to P. expansum species whereas the remaining strains (12.8%) belong to P. solitum. Furthermore, 67.7% of P. expansum strains produced patulin when grown on apple puree for 14 days at 25 °C with values ranging from 10.7 mg kg−1 to 125.9 mg kg−1, whereas all P. solitum did not produce the mycotoxin. This study highlights the presence of Penicillium spp. and their related mycotoxin risk during apple storage and calls for the implementation of proper measures to decrease the risk of mycotoxin contamination of apple fruit products.

Resumen El PVC es uno de los plásticos más usados y de los que más residuos se producen. En los últimos años, se han reportado microorganismos capaces de degradarlo y que en su mayoría provienen de ambientes en los que se acumula. El objetivo de este trabajo fue estudiar la degradación del PVC sin plastificante, a partir de hongos aislados de una muestra de suelo contaminada con resina de PVC. Los hongos se aislaron y caracterizaron morfológicamente, obteniendo 30 aislamientos de los que se escogieron 8 para someterlos a pruebas preliminares, en un medio cuya única fuente de carbono era una película de PVC. Se hicieron curvas de crecimiento de los dos aislamientos con mejores resultados, y su identificación molecular mostró que correspondían a Penicillium sp. y Mucor sp.. Este último ganó biomasa a partir del PVC y en los dos casos, las películas mostraron cambios visibles, que fueron respaldados por espectros de infrarrojo. Aunque los resultados mostrados en este artículo son preliminares, abren la puerta a nuevas formas de degradación de los residuos de PVC, que son muy persistentes. Abstract PVC is one of the most widely used plastics today and the one that produces the most waste. In recent years, microorganisms capable of degrading it have been reported, most of which come from environments in which PVC is accumulated. The aim of this work was to study the degradation of PVC without plasticizer, from fungi isolated from a soil sample contaminated with PVC resin. The fungi were isolated and morphologically characterized, 30 morphospecies were obtained, 8 were chosen to undergo preliminary tests in a medium whose only carbon source was a PVC film. Growth curves of the two isolates with better results were made and their molecular identification showed that they corresponded to Penicillium sp. and Mucor sp., the latter gained biomass from PVC and in both cases, the films showed visible changes, which were supported by infrared spectra. Although the results shown in this article are preliminary, they open the door to new forms of PVC waste degradation that are very persistent.

As a multifunctional signaling molecule, hydrogen sulfide (H2S) has been reported to induce plant responses to a variety of abiotic stresses. However, there are no reports on H2S treatment inducing resistance in apples against Penicillium expansum, a biotic factor, and its possible mechanism of action. In this study, fumigating apples with 5 mM sodium hydrosulfide (NaHS), the exogenous donor of H2S, for 12 h reduced the diameter of lesions in fruit colonized by P. expansum. NaHS treatment markedly promoted the synthesis of endogenous H2S, hydrogen peroxide (H2O2), and nitrogen oxide (NO). In vivo NaHS treatment enhanced the activities of phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, p-coumarate:coenzyme A ligase isoenzymes, caffeoyl-CoA-O-methyltransferase, caffeic acid-O-methyltransferase, ferulic acid-5-hydroxylase, cinnamyl-CoA reductase, and cinnamyl-alcohol dehydrogenase. The treatment also facilitated the production of specific phenolic acids, such as cinnamic acid, p-coumaric acid, caffeic acid, ferulic acid, and sinapic acid; total phenolic compounds; p-coumaryl alcohol; coniferyl alcohol; sinapyl alcohol; and lignin. NaHS treatment induced resistance against P. expansum in apples through H2O2- and NO-mediated activation of phenylpropanoid metabolism.

Author(s): Chamorro-Garcia, Raquel; Poupin, Nathalie; Tremblay-Franco, Marie; Canlet, Cecile; Egusquiza, Riann; Gautier, Roselyne; Jouanin, Isabelle; Shoucri, Bassem M; Blumberg, Bruce; Zalko, Daniel | Abstract: BackgroundEndocrine disrupting chemicals (EDCs) contribute to the etiology of metabolic disorders such as obesity, insulin resistance and hepatic dysfunction. Concern is growing about the consequences of perinatal EDC exposure on disease predisposition later in life. Metabolomics are promising approaches for studying long-term consequences of early life EDC exposure. These approaches allow for the identification and characterization of biomarkers of direct or ancestral exposures that could be diagnostic for individual susceptibility to disease and help to understand mechanisms through which EDCs act.ObjectivesWe sought to identify metabolomic fingerprints in mice ancestrally exposed to the model obesogen tributyltin (TBT), to assess whether metabolomics could discriminate potential trans-generational susceptibility to obesity and recognize metabolic pathways modulated by ancestral TBT exposure.MethodsWe used non-targeted 1H NMR metabolomic analyses of plasma and liver samples collected from male and female mice ancestrally exposed to TBT in two independent transgenerational experiments in which F3 and F4 males became obese when challenged with increased dietary fat.ResultsMetabolomics confirmed transgenerational obesogenic effects of environmentally relevant doses of TBT in F3 and F4 males, in two independent studies. Although females never became obese, their specific metabolomic fingerprint evidenced distinct transgenerational effects of TBT in female mice consistent with impaired capacity for liver biotransformation.DiscussionThis study is the first application of metabolomics to unveil the transgenerational effects of EDC exposure. Very early, significant changes in the plasma metabolome were observed in animals ancestrally exposed to TBT. These changes preceded the onset of obesogenic effects elicited by increased dietary fat in the TBT groups, and which ultimately resulted in significant changes in the liver metabolome. Development of metabolomic fingerprints could facilitate the identification of individuals carrying the signature of ancestral obesogen exposure that might increase their susceptibility to other risk factor such as increased dietary fat.

Penicillium expansum is an important postharvest pathogen of pomaceous fruit and a causal agent of blue mold or soft rot. In this study, we investigated the effect of ambient pH on growth, ultrastructure alteration, and pathogenicity of P. expansum, as well as accumulation of patulin and expression of genes involved in patulin biosynthesis. Under different pH, the fungus was routinely cultured and collected for growth, pathogenicity, patulin production, and gene expression studies using transmission electron microscopy, apple inoculation, HPLC, and RT-qPCR methods. Different ambient pH had significant impact on expression of genes and growth factors involved in patulin biosynthesis. Under same range of pH, gene expression profile, growth factors, and patulin accumulation (in vivo and in vitro) all showed similar changing trends. A well-developed cell was observed in addition to upregulation of genes at pH between pH 5.0 and 7.0, while the opposite was observed when pH was too basic (8.5) or too acid (2.5). Additionally, ambient pH had direct or indirect influence on expression of PecreaA, PelaeA, and PepacC. These findings will help in understanding the effect of ambient pH on growth, pathogenicity, and patulin production and support the development of successful methods for combating P. expansum infection on apple fruits.

Recently, the use of nanoparticles has been increased in improving plant seedlings. In this study, antifungal nanocomposite (St@CuONPs) based on starch and biosynthesized copper oxide nanoparticles (CuONPs) using Aspergillus niger AH1 was fabricated to improve rooting and seedlings of banana. CuONPs and St@CuONPs were fully characterized using attenuated total reflection-Fourier transform infrared (ATR-FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and transmission electron microscopy (TEM) techniques. CuONPs and St@CuONPs were evaluated as antifungal activity as well as to improve rooting and seedlings of banana. Results of antifungal activity revealed that CuONPs and St@CuONPs have promising antifungal activity toward Penicillium expansum, Fusarium oxysporum, and Rhizoctonia solani, where St@CuONPs was slightly higher than CuONPs. Meanwhile, different concentrations of CuSO4 and St@CuONPs were used to determine the effects on morphological growth parameters, e.g., leaf numbers, shoot, and root length of banana plantlets. The results showed that the highest increasing in the number of leaves (8.56 leaves), roots formed per explants, and root length (6.40 cm) was noticed in medium supplemented with 9 mg L–1 St@CuONPs. Also, the data revealed that the highest amount of chlorophyll was recorded with 9 mg L–1 St@CuONPs (33.87 SPAD values). In conclusion, St@CuONPs as nanocomposite is promising to improve rooting and seedlings of banana as well as to protect it from fungal plant diseases.

Nowadays, the extract of seaweeds has drawn attention as a rich source of bioactive metabolites. Seaweeds are known for their biologically active compounds whose antibacterial and antifungal activities have been documented. This research aimed to study the profile of phenolic compounds using the HPLC method and determine biologically active compounds using the GC-MS method and the antifungal activity of Gracilariopsis persica against plant pathogenic fungi. G. persica was collected from its natural habitat in Suru of Bandar Abbas, Iran, dried, and extracted by methanol. The quantitative results on phenolic compounds using the HPLC method showed that the most abundant compounds in G. persica were rosmarinic acid (20.9 ± 0.41 mg/kg DW) and quercetin (11.21 ± 0.20 mg/kg DW), and the least abundant was cinnamic acid (1.4 ± 0.10 mg/kg DW). The GC-MS chromatography revealed 50 peaks in the methanolic extract of G. persica, implying 50 compounds. The most abundant components included cholest-5-en-3-ol (3 beta) (27.64%), palmitic acid (17.11%), heptadecane (7.71%), and palmitic acid methyl ester (6.66%). The antifungal activity of different concentrations of the extract was determined in vitro. The results as to the effect of the alga extract at the rates of 200, 400, 600, 800, and 1000 μL on the mycelial growth of four important plant pathogenic fungi, including Botrytis cinerea, Aspergillus niger, Penicillium expansum, and Pyricularia oryzae, revealed that the mycelial growth of all four fungi was lower at higher concentrations of the alga extract. However, the extract concentration of 1000 μL completely inhibited their mycelial growth. The antifungal activity of this alga may be related to the phenolic compounds, e.g., rosmarinic acid and quercetin, as well as compounds such as palmitic acid, oleic acid, and other components identified using the GC-MS method whose antifungal effects have already been confirmed.

“Lianzhifan solution” (LZF) is produced by the natural fermentation of coptis root and gardenia fruit, and it is a classic prescription for external use in anorectal department. During the fermentation process, the structural evolution of microbial communities led to significant changes in the chemical profile. In this study, we first analyzed the dynamic changes of chemical components as well as the composition and succession of microbial community during the whole fermentation process of LZF, and confirmed the changes of characteristics of nine compounds during the whole fermentation process by metabolic profile. Further analysis found that there was no significant change of alkaloids in all stages of fermentation of LZF, but there were significant changes of iridoids in the middle and late stage of fermentation by deglycosylation. Genipin gentiobioside and geniposide were converted to genipin by biotransformation, showing that deglycosylation was the main event occurring in the fermentation. The community composition and abundance of species in 10 and 19days LZF fermentation broth were analyzed with high-throughput sequencing technology, and 16 dominant bacterial genera and 15 dominant fungal genera involved in the fermentation process were identified. Correlation analysis revealed that Penicillium expansum and Aspergillus niger involved in the fermentation were the dominant genera closely related to the dynamic changes of the deglycosylation of the main chemical components, and P. expansum YY-46 and A. niger YY-9 strains were obtained by the further fractionation. Then the monoculture fermentation process was evaluated, whereby we found that the deglycoside conversion rate of iridoid glycosides was greatly improved and the fermentation cycle was shortened by 3–4 times. This finding combined with equivalence evaluation of chemical component and pharmacodynamics to confirm that P. expansum YY-46 and A. niger YY-9 strains were key strains for fermentation concoction. This study established an efficient and practical screening strategy “Microfauna communities-Chemical component-Pharmacodynamic” axis for key strain, to improve the production process and formulating good manufacturing practice (GMP) work, and it is also applicable to the whole fermentation drugs industry.Graphical AbstractThe figure highly summarizes the research content of this study and shows the screening process of key strains in LZF fermentation.

The present study aimed to assess the antimicrobial properties of encapsulated lyophilized powdered extracts of pomegranate peels (PP), avocado peels (AP) and avocado seeds (AS) in vitro and in vivo. Minimum Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) methods, optical density measurement, and well diffusion assay were used to determine antimicrobial activity against food borne bacteria (Gram− Escherichia coli, Salmonella typhimurium, Campylobacter jejuni, Pseudomonas putida), (Gram+ Staphylococcus aureus, Listeria monocytogenes, Clostridium perfringens, Lactobacillus plantarum), and fungi (Penicillium expansum and Aspergillus niger) based on a mixture design model. Additionally, the most effective powder was studied in vivo in yogurt, cream cheese, and minced meat burger. The samples that contained high polyphenol content also exhibited higher antioxidant, antimicrobial, and antifungal activity. From the results of the well diffusion, the MIC/MBC, and the cell optical density assays, the antimicrobial activity of the extracts was found to be correlated to the total phenolic content (TPC) of the samples and the type of the microorganism. The pomegranate peels extract presented the higher TPC and antioxidant activity and constitute the highest percentage in the most active antimicrobial mixture. The powders that were tested in vitro showed microbial type-dependent effects in each food model. The results presented here can be further studied in the large-scale industrial production of natural food preservatives.

Penicillium expansum, as the causal agent of blue mould and a main producer of mycotoxin patulin, is a global concern for economic and food safety. To date, the nutritional requirements of the pathogen during infection and patulin biosynthesis are poorly understood. Here, we genetically characterized the role of the bZIP transcription factor PeMetR in sulfur metabolism, virulence and patulin biosynthesis of P. expansum. The PeMetR regulator is crucial for normal germination and growth on inorganic S-sources but dispensable for utilization of organic S-sources. Accordingly, it is involved in regulating the expression of genes in sulfur assimilation pathway rather than methionine metabolic processes. Disruption of PeMetR resulted in a complete loss of virulence on various fruits. Additionally, the mutant showed a remarkably reduced ability to produce patulin. Exogenous methionine could partially or completely rescue the impaired phenotypes of the mutant. Inactivation of the sulfur assimilation pathway genes, PesA, PesB, PesC, PesF, generated growth, virulence and patulin production defects similar to those of ΔPeMetR. Overall, our study provides evidence that PeMetR-mediated sulfur assimilation is essential for growth and infection and shows for the first time that regulation of sulfur assimilation affects biosynthesis of an important mycotoxin patulin in P. expansum.

To study the mechanism by which Pichia anomala induced with chitosan (1% w/v) controls blue mold disease in table grapes caused by Penicillium expansum, this study evaluated alterations in three yeast enzymatic activities. The changes in the five primary disease defense-related enzymes and two non-enzyme activities of table grapes were assayed. The results of the study showed that chitosan (1% w/v) significantly increased the yeast β-1,3-glucanase, catalase (CAT), and malondialdehyde (MDA) activities. Furthermore, P. anomala alone or induced with chitosan (1% w/v) significantly increased the table grapes enzymatic activities of Polyphenol oxidase (PPO), phenylalanine (PAL), peroxidase (POD), and catalase (CAT) compared to the control. The RT-qPCR results also confirmed that the genes of these major disease defense enzymes were up-regulated when the table grapes were treated with P. anomala. The highest results were recorded when the fruit was treated by yeast induced with chitosan (1% w/v). The phenolic compounds, in addition to their nutritional value, can also increase the antimicrobial properties of table grapes. The current experiment determined that the total phenol and flavonoid contents of table grapes showed the highest results for fruits treated by P. anomala induced with chitosan compared with the control. Generally, the increment of these fruit enzymatic and non-enzymatic activities shows improved table grape defense against the pathogenic fungus. The induction of the yeast with chitosan also increases its bio-control efficacy against the pathogen. This study will enable future detailed investigation in the yeast pathogen control mechanisms and the use of yeasts as bio-pesticides.

Several economically important crops, fruits and vegetables are susceptible to infection by pathogenic fungi and/or bacteria postharvest or in field. Recently, plant essential oils (EOs) extracted from different medicinal and officinal plants have had promising antimicrobial effects against phytopathogens. In the present study, the potential microbicide activity of Mentha × piperita cv. ‘Kristinka’ (peppermint) EO and its main constituents have been evaluated against some common phytopathogens. In addition, the cell membrane permeability of the tested fungi and the minimum fungicidal concentrations were measured. The antifungal activity was tested against the following postharvest fungi: Botrytis cinerea, Monilinia fructicola, Penicillium expansum and Aspergillus niger, whereas antibacterial activity was evaluated against Clavibacter michiganensis, Xanthomonas campestris, Pseudomonas savastanoi and P. syringae pv. phaseolicola. The chemical analysis has been carried out using GC-MS and the main components were identified as menthol (70.08%) and menthone (14.49%) followed by limonene (4.32%), menthyl acetate (3.76%) and β-caryophyllene (2.96%). The results show that the tested EO has promising antifungal activity against all tested fungi, whereas they demonstrated only a moderate antibacterial effect against some of the tested bacteria.

The use of natural compounds as food preservatives is becoming increasingly popular as it is perceived positively by consumers. Among these substances, essential oils have attracted great interest owing to their antioxidant and antimicrobial properties. However, several challenges impair the use of essential oils in food products, such as their degradation or loss during food processing and storage, the strong aroma, even at low concentrations, which may negatively affect the sensory characteristics of food. In this context, the development of nanoformulations able to stabilize essential oils may represent a smart solution to this issue. The aim of the study was to evaluate the efficiency of alginate-based nanoformulations enriched with lemongrass (Cymbopogon nardus) essential oil (LEO) and Tween 80 against several fungi namely Penicillium expansus, Aspergillus niger and Rhizopus spp. Firstly, the flow behavior of systems at different concentrations of alginate (1%, 2% and 3% w/w) were studied. Then, emulsion-based nanoformulations at different concentrations of lemongrass essential oil in the range of 0–2% w/w were stabilized by a fixed amount of Tween 80, characterized and tested for their antifungal activity. Our results showed that the best nanoformulation able to inhibit Rhizopus spp., Penicillium expansum and Aspergillus niger, for at least 10 days, was constituted by 1% alginate/1.5% LEO/1% Tween 80. Hence, the incorporation of essential oil into nanoformulation systems may represent a valid alternative to overcome the disadvantages that limit the commercial application of essential oils.

AIMS This research sought to improve the ability of biocontrol yeast to suppress postharvest fungal disease and explore possible mechanisms of action. METHODS AND RESULTS The addition of 2% sodium glutamate (SG), which is edible and recognized as safe, enhances the inhibitory effect of Rhodosporidium paludigenum Fell & Tallman on Penicillium expansum in vivo and in vitro. Rhodosporidium paludigenum cells grown in medium with a final concentration of 2% SG, displayed viability under a variety of stress conditions, including sodium chloride (NaCl), calcofluor white (CFW), Congo red (CR) and sodium dodecyl sulphate (SDS). Activity and relative gene expression levels of antioxidant-related enzymes in R. paludigenum, including peroxisomal catalase (CAT), thioredoxin reductase (TrxR), glutathione peroxidase (GSH-PX), glutathione reductase (GR) and superoxide dismutase (SOD) were altered in the presence of SG. Levels of reactive oxygen species (ROS) increased in cells grown in the presence of SG as well as the content of several amino acids. CONCLUSIONS In the presence of 2% SG R. paludigenum inhibited P. expansum and exhibited tolerance to a number of stressful conditions which may involve the upregulation of antioxidant enzymes and amino acids. SIGNIFICANCE AND IMPACT OF THE STUDY The ability of culture conditions to enhance the fungal suppressive abilities of yeast has the potential to enhance the management of postharvest disease in fruit.

Recent apple-related recall and outbreak events have exposed a need for better food safety controls along the supply chain. Following harvest, apples can be stored under a controlled atmosphere for up to 1 year after harvest before packing and distribution, making the crop susceptible to many opportunities for contamination that increase the quantity of postharvest losses. Botrytis cinerea and Penicillium expansum cause significant rot-associated losses to the apple industry. These fungi can colonize and destroy apple tissue as storage duration increases, which may also impact the growth of saprophytic foodborne pathogens like Listeria monocytogenes. Thus, the objective of this study was to observe population changes of Listeria innocua as a surrogate for L. monocytogenes on apples inoculated with B. cinerea or P. expansum under long-term controlled atmosphere cold storage conditions to identify the effect of postharvest mold growth on growth patterns of a microorganism relevant to food safety. 'Gala' and 'WA 38' apples (n = 1,080) were harvested, treated with pyrimethanil, and inoculated with L. innocua only or with L. innocua and one of the mold species on wounded and unwounded portions of the apple equator. Apples were treated with 1-methylcyclopropene and stored at a controlled atmosphere (2 kPa O2, 1 kPa CO2, 1°C) for 1 week and 1, 3, 6, 9, and 11 months before enumeration. After 3 months, L. innocua consistently fell below the limit of detection (2.35 Log CFU/g), and samples were enriched following a modified Bacteriological Analytical Manual method with PCR confirmation. Listeria persistence was dependent on the storage duration and type of fungal contamination (P ≤ 0.05). Surface wounding may impact these trends, depending on the apple variety. Prevalence of L. innocua was greater in Gala apples. Future studies should more closely examine the interactions on the fruit surface that occur during the seemingly critical time frame of 3 to 6 months in storage.