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Mycoremediation is an important process that targets the removal of petroleum hydrocarbons by fungi. Fungi have advantages with their extensive enzymatic systems, rapid adaptation to toxic organic pollutants, and to adverse environmental conditions. In this study, the colorimetric method was used for the preliminary investigation of petroleum degradation with ten fungal strains. Petroleum degradation ability of spore suspension, live biomass (fungal pellet and disc) and cell-free culture supernatant of the potent A. ochraceus strain were investigated by gravimetric analysis. It was found that the fungal disc (94%) was more successful than the spore suspension (87%) in petroleum degradation under physiological conditions determined as pH:5.0, 1% of petroleum concentration, 5% (v/v) of inoculum concentration (with spore suspension) and 1 g/100 mL of inoculum amount (with fungal disc) and 7 days of the incubation period. The degradation rate constant and half-life period of spore suspension were calculated as 0.291 day−1 and t1/2 = 0.340 and of the fungal disc were 0.401 day−1 and t1/2 = 0.247. Although, 7.5% and 10% (v/v) concentration of cell-free culture supernatant achieved more than 80% petroleum removal, it was not as effective as a fungal disc. According to gas chromatography/mass spectrometry analysis, the fungal disc of A. ochraceus strain degraded long-chain n-alkanes such as C35 and C36 more effectively than n-alkanes in the range of C22–C34. The fact that the A. ochraceus NRRL 3174 strain has a high petroleum degradation capacity as well as being a potent biosurfactant producer will provide a different perspective to advanced mycoremediation studies.

Podang mango, an endemic mango from Kediri, has a short post-harvest shelf life that accelerates by physiological factors and pathogenic fungal contamination. Chitosan reported as an active compound for coating. The apple-snail was reported to contain chitosan in its shell. The purposes of this study are to synthesis chitosan from Pila ampullacea, examine anti-fungal activities to Fusarium oxysporum and Aspergillus ochraceus, and verify the effect of chitosan coating on shelf life and quality of Podang mango. The effect of chitosan coating on fruit quality was analyzed using three treatments which were negative control, chitosan, and chitosan-gelatin with 3 replications. The parameters measured were color, texture, aroma, water content, vitamin C, and weight loss. Results showed that chitosan synthesized from P. ampullacea had 53.56 degrees of deacetylation. Antifungal activity test of chitosan 2% had the highest inhibitory activity against F.oxysporum and A.ochraceus growth with 14.46 mm and 10.33 mm inhibition zone. Coating with chitosan statistically affected fruit quality for 14 days of storage (sig

The present study describes the synthesis, characterization and antiochratoxin efficacy of zinc oxide nanoparticles against hepatotoxicity and nephrotoxicity in albino rats. Ochratoxin A (OTA) was produced by solid state fermentation on wheat grains using Aspergillus ochraceus and estimated by HPLC. ZnONPs were produced using leaf extract of Camellia sinensis and characterized by UV–visible spectrophotometer, Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR) and X-ray diffraction (XRD). Different doses were given to albino rats by dividing them into different groups i.e., group A (basal feed), group B (50 ppb OTA), group C (50 ppb OTA and 25 ppb ZnONPs) and group D (25 ppb ZnONPs). Blood was collected for biochemical analysis and tissues of liver and kidney were taken for histopathological examination. UV–visible spectroscopy showed prominent peak of green synthesized nanoparticles having maximum absorption of 350 nm and size range from 30 to 40 nm using SEM. The synthesized ZnO nanostructure was confirmed by XRD image and different functional groups responsible for green synthesis were confirmed by FTIR analysis. OTA significantly decreased the weight gain, feed consumption and feed conversion ratio of rats. Histopathological examination of liver and kidney indicated that OTA has severe effects including degeneration, increase sinusoidal spaces, infiltration of lymphocytes, vascular congestion and necrosis. Biochemical parameters such as ALT, AST and creatinine were significantly raised whereas albumin and total protein were decreased by OTA feeding. These changes were ameliorated by green synthesized ZnONPs. This study concluded that OTA severely affect liver and kidney of rats and green tea mediated ZnONPs plays a hepatoprotective and nephroprotective role to reduce the lethal effects of OTA in albino rats.

The principle aim of this work is to evaluate the antimicrobial and antiochratoxic power of the essential oil of Mentha spicata (EOM). This oil was obtained by hydrodistillation with a yield of 0.3%. EOM was screened for its possible antibacterial activity in vitro against five strains of pathogenic bacteria, using the solid disc diffusion method and the microdilution method. The extract reacted positively to the bacterial strains tested. The results of the antifungal activity show an inhibition of mycelial growth; it is total on Aspergillus ochraceus, Aspergillus flavus, Aspergillus niger, Penicillium viridicatum, and on Rhizopus stolonifer and Alternaria alternate at a concentration of 12.5 μl/ml. Tests carried out on yeasts mainly referenced as Candida albicans IP444 and Candida albicans 2679 showed minimal inhibition (MIC) of the order of 7.81 μl/ml and 0.97 μl/ml, respectively and theMICis of the order of 0.48 μl/ml for Candida albicans 10 231. In addition, the EOM antiochratoxigenic test for ochratoxin A production capacity following a series of high performance liquid chromatography separation and ultraviolet detection (HPLC–UV) analyses detected an inhibitory effect proportional to the EOM concentration. In general, the essential oil showed an effective antimicrobial action on the growth of the tested pathogens and an antiochratoxigenic.

Seventy-five samples of selected meat products, including luncheon, beef burger, sausage, basterma, and kofta, were collected from Alexandria and New Borg El-Arab cities (Egypt). The samples were subjected to mycological examination as well as for detection of aflatoxin B1 (AFB1) and ochratoxin A (OTA) residues. Besides, the study evaluated the effect of aqueous leaf extracts from mealycup sage (Salvia farinacea) and neem (Azadirachta indica), individually and in combination, on the growth of human pathogens Aspergillus parasiticus and Aspergillus flavus producing AFB1, as well as Aspergillus ochraceus and Aspergillus niger which produce OTA. The obtained results revealed that sausage samples had the highest mould count with a mean value of 13.20×102/g, followed by basterma samples 12.05×102/g, then beef burger 7.39×102/g. In contrast, luncheon and kofta samples had the lowest count with a mean value of 5.51×102/g and 2.82×102/g. The findings revealed the antifungal potential of tested extracts. The total inhibition of A. parasitcus and A. niger growth was observed at 2 mg/mL of the combined extract. Salvia farinacea extract had the highest total phenolic content and total flavonoid content with a value of 174.1 and 52.6 mg g-1, respectively. Rutin was the major phenolic component in neem and combined extracts, accounting for 19123 and 8882 μg/g, respectively. Besides, the study investigated detoxification of AFB1 and OTA using combined extract in albino rats. The results confirmed the convenient and safe use of Salvia farinacea and Azadirachta indica extract and their combination as natural antifungal and antioxidant agents. The combined extract could be used as a natural preservative in food processing to control or prevent contamination.

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%.

The growing demand and application of industrially important enzyme necessitate the need to explore new sources with diverse enzymes ranging in their specificity and activities. Enzymes are safe alternatives to chemical synthesis due to minimum side effect and ease of manufacturing. Solid state fermentation (SSF) is a cost-effective alternative to submerged fermentation with agro-residues or waste, often being used as substrate for growing diverse organisms for production of metabolites. Current study is one of the scarce report on exploring alpha amylase and cellulase production ability Aspergillus ochraceus (MTCC 1877) using wheat bran as substrate at relative humidity of 90% and at 30 ºC, for 7 days. Result showed the potential of Aspergillus ochraceus (MTCC 1877), as potential source of the two enzymes. Results revealed comparatively higher alpha amylase activity in the SSF extract of Aspergillus ochraceus (MTCC 1877) in comparison to Trichoderma longibrachiatum (ITCC 7839). On the contrary, comparatively higher cellulase activity was observed in the SSF extract of Trichoderma longibrachiatum (ITCC 7839). The results showed the potential of Aspergillus ochraceus (MTCC 1877) as a source of the two enzymes. Variation in enzymes activity may be attributed to the experimental culture conditions and may be further optimized to enhance the enzymes yield.

The search for natural sources to control microorganisms is of interest in food production. This study evaluated the chemical composition and antimicrobial activity of the essential oil from Psidium cattleianum leaves. The essential oil was extracted by hydrodistillation, and identified by GC-MS. The predominant class of compounds was sesquiterpenes (47.6%) and the major compounds were trans-β-caryophyllene (14.7%), 1,8-cineole (11.7%) and ɣ-muurolene (5.6%). The antimicrobial activity was carried out by microdillution technique against eight fungi and eight bacteria. The minimum inhibitory concentration ranged from 0.17 to 11.25 mg mL-1 for fungi, and from 1.40 to 16.87 mg mL-1 for bacteria. The highest activities were against fungi Aspergillus fumigatus (ATCC 1022), Aspergillus ochraceus (ATCC 12066), Aspergillus versicolor (ATCC 11730), and Trichoderma viride (IAM 5061), and bacteria Pseudomonas aeruginosa (ATCC 27853), Bacillus cereus (clinical isolate), and Staphylococcus aureus (ATCC 6538) with potential to prevent foodborne diseases.

Ochratoxin A (OTA) is a mycotoxin produced by several fungal species including Aspergillus ochraceus, A. carbonarius, A. niger and Penicillium verrucosum.

Citrinin (CIT), a known nephrotoxic mycotoxin, is mainly produced by Penicillium, Aspergillus, and Monascus species. It is a natural contaminant in cereal grains, foods, and feedstuff. Liupao tea (or Liubao tea) is a typical Chinese dark tea obtained via indigenous tea fermentation facilitated by microorganisms. Certain fungi present in Liupao tea that may produce CIT are a potential threat to consumer health. In the present study, various potential toxigenic mycoflora and the natural occurrence of CIT in Liupao tea were surveyed via the culture-dependent method, high performance liquid chromatography-fluorescence detection (HPLC-FLD), and ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Total mold counts ranged from 3.5 × 102 CFU/g to 2.1 × 106 CFU/g tea in 28 tea samples. A total of 218 fungal isolates belonging to five genera and 23 species were identified. Some of these strains, such as Aspergillus ochraceus, Aspergillus oryzae, Penicillium citrinum, and Penicillium chrysogenum, may potentially be a CIT-producing species. In addition, 32.7% of 113 Liupao tea samples were contaminated with CIT at concentrations ranging from 7.8 to 206.1 μg/kg. These CIT concentrations in Liupao tea are chiefly attributed to climatic conditions and water activity during storage that favor fungal proliferation and mycotoxin production. However, CIT could not be detected in Liupao tea stored for over 10 years. These results provide the first information about the potential toxigenic mycoflora and natural occurrence of CIT in Liupao tea. Therefore, storage conditions and fungal community must be monitored to ensure the quality of Liupao tea.

Aspergillus ochraceus is a soil fungus known to produce ochratoxin A, a harmful secondary metabolite. Prevention and control of fungal pathogens mostly rely on chemical fungicides, which is one of the contributing factors in the emergence of the fungal resistance, hence novel methods for fungal eradication have been extensively researched. The cold atmospheric pressure (CAP) plasma generated in ambient air has been recently applied in microbial decontamination. Here we used the diffuse coplanar surface barrier discharge in inactivation of a toxigenic strain A. ochraceus. The plasma-treated conidia and mycelium exhibited morphological changes such as ruptures and desiccation. Mycelium dehydration and changes in the chemical composition of hyphal surface accompanied plasma treatment. The growth of 26 h old mycelia were significantly restricted after 30 s of plasma treatment. The conidial vitality declined 4 logs after 180 s of plasma exposure leading to almost complete decontamination. After shorter plasma treatment of conidia, the ochratoxin A (OTA) production increased at the early stage of cultivation, but the overall level was significantly reduced compared to untreated samples after longer cultivation. Our results indicated that the fungal growth and the OTA production were significantly changed by plasma treatment and underscored CAP plasma as a promising method in the decontamination of A. ochraceus without a risk to generate strains with increased OTA production.

Artemisia is the largest and most diverse genus from the Asteraceae family that named locally "Dermaneh" in Iran. This study was conducted to determine, the chemical compounds of Artemisia persica Boiss essential oil and its antifungal effect, toward six toxigenic fungal strains in vitro. The yield of essential oil from the aerial parts of this plant species, using hydrodistillation method obtained 0.18% (v/w). The results of GC/MS analysis identified 31 components in the essential oil that laciniata furanone E (17.1%), artedouglasia oxide C (13.2%), Trans-pinocarveol (10.2%), pinocarvone (8.5%), and α-pinene (5.8%) were the major compounds. The results of the antifungal activity showed that the most sensitive fungal strains to A. persica Boiss. essential oil were Aspergillus ochraceus and Aspergillus parasiticus with lower minimal fungicidal concentration (MFC) of 1.25 μl/ml (v/v). Also the strong fungicidal effect was observed against Aspergillus flavus and Aspergillus nidulans at a MFC value of 2.5 μl/ml, while the fungicidal activity against Aspergillus fumigatus and Aspergillus niger observed in the 10 μl/ml oil concentration. According to the results A. persica Boiss essential oil has a acceptable antifungal activity against Aspergillus strains and can be used to prevent food crops from fungal contaminations.

Essential oils can be considered antifungal agents in controlling food contamination, due to their biological potential. Eremanthus erythropappus essential oil (EEO) was extracted by hydrodistillation, and its chemical composition was evaluated by gas chromatography using mass spectrometer (GC‐MS) and flame ionization detectors (GC‐FID). EEO antifungal and antimycotoxygenic effect was evaluated on different species of Aspegillus genus. The main component of the EEO was α‐bisabolol (98.853%). The essential oil exhibited fungicidal activity at concentrations of 0.98, 15.63, and 15.63 μL/mL against Aspergillus carbonarius, Aspergillus flavus, and Aspergillus ochraceus, respectively. The application of the EEO significantly inhibited ochratoxin A production that ranged from 62.06% to 84.27% and from 68.29% to 98.44% for the fungal species A carbonarius and A ochraceus, respectively, and also inhibited the production qualitatively of aflatoxins B₁ and B₂. Ergosterol biosynthesis of fungi A carbonarius (76.56%‐92.07%), A flavus (81.28%‐97.75%), and A ochraceus (47.61%‐85.56%) significantly decreased in the presence of EEO. Electromicrographs showed morphological alterations and some damages caused by EEO in the fungal membranes of three fungi species. There was a change in the hyphae uniformity present in the fungi and also loss of conidia integrity in A carbonarius, and no conidia formation in A flavus and A ochraceus, after treatment with the essential oil. Therefore, the results suggested that EEO was capable of inhibit fungal growth, ergosterol biosynthesis, and the production of mycotoxins that can contaminate food.

The present study aims to investigating, the chemical composition of the essential oil extracted from Thymus capitatus harvested in west, Algeria. It seeks the other objective to the determination of antioxidant, antibacterial and antifungal activities of oil in order to find new metabolite products, which are characterized by a biological activity. The investigations and research was carried out on the essential oil (EO) extracted from the dried leaves of Thymus Capitatus Lamiaceae that was harvested in the region of Tiart (West of Algeria). GC–MS analysis of EO is identified 22 components while thymol, carvacrol and γ-terpinene are the major components. The antioxidant activity of EO was evaluated by three methods: the DPPH test (2,2-diphenyl-1-1-picrirylhydrazil), the FRAP (Ferric reducing antioxidant power) and TAC (Total Antioxidant Capacity) test and compared with ascorbic acid, BHA (Butylated hydroxy anisole) and commercial thymol as standards. The results indicated that Thymus Capitatus oil showed higher antioxidant power in comparison with the standards with values of (0.61, 2.13 and 0.78) μg/mL for the DPPH, FRAP and TAC tests respectively. Also, the antibacterial activity of the oil was tested using the agar disc diffusion method, by determining the inhibition zone and the minimum inhibitory concentration (MIC). In this case, the results have shown a potential of the antibacterial activity than commercial antibiotics (Chloramphenicol, Cefoxitin, Gentamycin) against the tested strains, with MIC values of (29.44, 14.72, 14.72 and 7.36 μg/mL) for Staphylococcus aureus, S. typhi, E. coli and Streptococcus pneumoniae strains respectively. Furthermore, the antifungal study on the fungi of Cladosporium herbarum, Alternaria infectoria, Aspergillus Ochraceus, Trichophyton Sp reported significant results. The effectiveness of the oil showed that the essential oil has a fungicidal effect against the strains Cladosporium herbarum, Alternaria infectoria and a fungistatic effect against strains Aspergillus ochraceus and Trichophyton rubrum. The research outcomes clearly demonstrate that the essential oils of thymus capitatus can present an interesting alternative naturel which can be useful for food preservation and pharmaceutical treatment.

Essential oils can be a useful alternative to the use of synthetic fungicides because they have biological potential and are relatively safe for food and agricultural products. The objectives of the present study were to evaluate the antifungal and antimycotoxigenic activities of the essential oils from Satureja montana L., Myristica fragrans H. and Cymbopogon flexuosus S. against Aspergillus flavus and Aspergillus ochraceus, as well as their effects on ergosterol synthesis and membrane morphology. The antifungal potential was evaluated by mycelial growth analysis and scanning electron microscopy. Fungicidal effects against A. flavus, with MFC of 0.98, 15.62 and 0.98 µL/mL, respectively, were observed for the essential oils from S. montana, M. fragrans and C. flexuosus. Aspergillus ochraceus did not grow in the presence of concentrations of 3.91, 15.62 and 0.98 µL/mL of the essential oils from S. montana, M. fragrans and C. flexuosus, respectively. The essential oils significantly inhibited the production of ochratoxin A by the fungus A. ochraceus. The essential oils also inhibited the production of aflatoxin B1 and aflatoxin B2. The biosynthesis of ergosterol was inhibited by the applied treatments. Biological activity in the fungal cell membrane was observed in the presence of essential oils, given that deleterious effects on the morphologies of the fungi were detected. The essential oils under study are promising as food preservatives because they significantly inhibit toxigenic fungi that contaminate food. In addition, the essential oils hindered the biosynthesis of mycotoxins.

The kiwi fruit has been drawing attention and a great deal of interest because of its health benefits. It is consumed in its natural form, while it is being presented in processed form by the food industry such as sweets, ice creams, frozen juice or pulp, and many other byproducts. The peel of kiwi fruit which is a byproduct of fruit is still under exploration, but it has raised much interest in this by-product because this has many bioactive molecule contents in it such as phenolic compounds. Kiwi fruit has shown antimicrobial activity apart from the antioxidant activity against many pathogenic bacteria i.e. Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes as well as fungi like Penicillium funiculosum, Aspergillus niger, Aspergillus versicolor and Aspergillus ochraceus. With the presence of antibacterial, antifungal, and antioxidant activities in kiwi fruit, it may be used as a potential medicinal fruit.

The antibacterial effect and mechanism of ozone (O3) treatment on freshly-peeled garlic inoculated with Gibberella intermedia (G. intermedia) and Aspergillus ochraceus (A. ochraceus) in different conditions were studied in vivo. The results showed that the specific O3 concentration, treatment time, and humidity significantly affected the garlic's indexes. The optimal treatment conditions of garlic inoculated with both molds were 6 ppm, 15 min, and 90%, respectively. After treatment with those conditions, the incidence etc of garlic were significantly reduced during storage. Differential analysis was performed for the RNA-sequencing and gene expression of the molds subjected to O3 treatment, as well as the samples that were not. The results showed that O3 treatment inhibited the growth of G. intermedia and A. ochraceus. Consequently, 2754 and 2378 differentially expressed genes, 1456 and 1591 up-regulated genes, and 1298 and 787 down-regulated genes were screened, respectively, for these molds, affecting the catalytic activity and various other pathways.

The occurrence of ochratoxin A (OTA) in matured meat products can be attributed to mycotoxin coming from raw materials and/or molds that develop on the product surface during ripening. This work aimed to evaluate OTA production by Aspergillus westerdijkiae inoculated on the surface of sausages and its diffusion into the product throughout ripening, study the effect of relative humidity (RH) on the production of this mycotoxin, and investigate the presence of OTA in dry fermented sausages that naturally present intense contamination by A. westerdijkiae. Italian-type sausages were surface inoculated with A. westerdijkiae and two distinct experiments were performed. In the first, the sausages were matured in a chamber with decreasing RH (from 95 to 75%) for 35 days. In the other, the sausages were incubated under different RH (79, 85 or 95%) for 21 days. Samples were taken at the beginning of the experiments and every 7 days, subdividing into casing, outer border, and core for analyses. Sausage samples naturally spoiled by ochratoxigenic fungi were collected during sanitary inspection. Even in the presence of A. westerdijkiae mycelia, no OTA was detected for up to 7 days of sausage maturation. On the other hand, this study demonstrated that the growth of A. westerdijkiae on salami surface produces high amounts of OTA on the casing and allows its diffusion through the casing with contamination to the outer border of sausages. In the same way, it shows that under similar water activity values of substrate, RH influences the amount of OTA produced. Conversely, OTA was restricted to the casing in the naturally contaminated sausages.

A total of 130 samples of ready-to- eat beef products comprised luncheon, fried sausage, fried burger, grilled kofta and fried liver (26 each) were randomly purchased embedded in their sandwiches ( with exception of luncheon's samples ) from different supermarkets and restaurants having various sanitation levels at Mansoura city, Egypt –were analysed mycologically. Laboratory findings exhibited presence of moulds contaminants in 23 (88.5 %) beef luncheon, 19 (73.1 %) every of fried beef sausage and fried beef burger, 18 (69.2 %) of grilled beef kofta plus 25 (96.2 %) fried beef liver samples. Whereas, the yeasts were detected in 25 (96.2%) luncheon, 21 (80.7%) sausage, 23 (88.5 %) burger, 22 (84.6%) kofta and 16 (61.5%) liver of the same samples. Enumeration plates of dichloran rose bengal chloramphenicol ( DRBC ) agar revealed the fungal populations in examined cooked beef products by maximum & mean ± standard error levels every of moulds and yeasts in 26 tissue samples of each product as 1.3×103 & 1.7 × 102 ± 0.48 × 102 and 6.5×103 & 1.6 ×103 ± 0.34 ×103 cfu / g in beef luncheon, 1.1×103 &1.6×102 ± 0.46×102 and 1.4×104 & 3 ×103 ± 0.67 × 103 cfu / g in fried beef sausage, 5×102 &1.1 × 102 ± 0.22 × 102 and 7.8 × 103 & 2.02 × 103 ± 0.48 × 103 cfu / g in fried beef burger, 1.2 × 103 & 2.4 × 102 ± 0.65 × 102 and 8.2 ×103 & 2.15 × 103 ± 0.51×103 cfu / g in grilled beef kofta plus 9.5 × 102 & 2.3 × 102 ± 0.48 × 102 and 6 × 103 & 8.92 × 102 ± 3.09 ×102 cfu / g in fried liver, respectively. Data revealed the isolation of a sum of 472 mould strains from a whole 130 tissue samples of cooked beef products ( 26 each of kind ) ; 88 (18.64 %) strains of them recovered from luncheon, 85 (18 % ) from fried sausage, 57 (12.08 %) from fried burger, 124 (26.27%) from grilled kofta, and 118 (25%) from fried liver samples. Generic level identification of the total mould strains categorized them as 170 (36.02 %) Aspergillus strains, 103 (21.82 %) Cladosporium, 76 (16.1%) Penicillium, 40 (8.47 %) Alternaria alternata and 22 (4.66 %) Fusarium strains which obtained from all kinds of tested beef products, in addition to 20 (4.24 %) Acremonium strictum strains (18 from fried sausage and 2 from grilled kofta), 16 (3.39 %) Geotrichum candidum (10 from fried sausage, 4 from fried burger and 2 from fried liver), 5 (1.06 %) Mucor (2 from luncheon and 3 from grilled kofta ), 4 ( 0.85 % ) Rhizopus ( 1 from luncheon, 2 from grilled kofta and 1 from fried liver ), 3 ( 0.64 %) Rhizomucor ( from grilled kofta samples), 2 (0.42%) each of Curvularia lunata (from grilled kofta and fried liver ) and Trichothecium roseum ( from fried burger ), 1 ( 0.21 % ) each of Bipolaris ( from fried liver ), Cheatomium ( from fried burger ), Drechslera ( from one grilled kofta ), Endomyces fibuliger ( from fried burger ), Ulocladium botrytis ( from grilled kofta ), besides 4 (0.85%) unidentified mould strains ( 3 from luncheon and 1 from fried liver samples ). Data showed the identified groups of total obtained 170 Aspergillus strains as 73 (42.94 %) of them were belonged to Aspergillus niger which were distributed in all types of tested samples and represented by 5 ( 6.85 % ) strains in luncheon, 3 ( 4.11 % ) in fried sausage, 10 (13.69 % ) in fried burger, 30 (41.09 %) in grilled kofta and 25 (34.25 %) in liver samples; followed by 23 (13.52 %) strains of Aspergillus fumigatus group consisted of 2 ( 8.69 %) strains recovered from luncheon, 10 ( 43.48 % ) from every of fried sausage and grilled kofta, besides 1 ( 4.35 % ) from fried burger ; then 22 (12.94 % ) strains of Aspergillus flavus group, 3 ( 13.64 %) of them isolated from luncheon, 4 (18.18 %) from grilled kofta and 15 ( 68.18 %) strains from fried liver samples ; besides 17 (10 %) isolates of Aspergillus ochraceus group, categorized as 6 ( 35.29 %) strains determined in each of luncheon & grilled kofta, 1 ( 5.88 %) in fried burger and 4 (23.53 %) in liver ; 10 (5.88 %) strains of Aspergillus terreus group were found in grilled kofta, exclusively ; 9 (5.29 %) isolates of Eurotium chevalieri, composed of 1 (11.11%) strains from every of luncheon & fried sausage, 3 ( 33.33 %) from grilled kofta, and 4 (44.44 %) strains from fried liver samples ; 6 ( 3.52 % ) strains of Aspergillus parasiticus, distributed as 1 (16.67 % ) of them recovered from each of fried sausage & fried liver and 2 ( 33.33% ) from every of fried burger & grilled kofta samples ; 5 (2.94%) isolates of Aspergillus candidus, 3 (60 %) of them were found in grilled kofta and 2 (40 %) in fried liver ; 4 ( 2.35 % ) strains of Eurotium amstelodami, consisted of 1 ( 25 % ) strain recovered from every of luncheon & grilled kofta and 2 ( 50 % ) from fried sausage samples ; in addition to a sole strain of Aspergillus ustus ( 0.58 % ) was found in a grilled kofta sample..

This study was aimed to find out an alternative to synthetic fungicides used in the control of fungi development in crops including coffee. The antifungal was extracted from green berries of Solanum indicum L. and its effect on the behavior of three Aspergillus species Ochratoxin A (OTA) producers (Aspergillus niger, Aspergillus carbonarius and Aspergillus ochraceus) was evaluated on a coffee-agar medium. The results show a reduction of growth with the increase of the antifungal content in the medium. Indeed, on the medium without antifungal, the growth rate has reached 90 mm within only 3 days of incubation for the three strains tested, while no growth was observed for A. niger and A.carbonarius from 8% of antifungal in the medium and for A. ochraceus, at 10% of antifungal in the medium even after 7 days of incubation. However, the total inhibition of OTA was observed from 8% of antifungal in the medium for these three strains of Aspergillus. Indeed, the amounts of OTA produced on the coffee-agar medium without antifungal were 0.005, 1.63 and 0.108 µg/mL respectively for A. niger, A. carbonarius and A. ochraceus, while from 8% of antifungal fraction in the medium, no detectable OTA amount was observed for the three strains tested after 7 days of incubation. The phytochemical screening of this antifungal fraction revealed compounds including flavonoids, carotenoids and saponins. These results suggest the use of this antifungal fraction as an alternative to synthetic fungicides for the control of fungi development and OTA production in coffee beans.

Introduction: Mycotoxin producing fungi are major contributors to food contamination and many epidemics in humans and animals. The adverse effects of the use of chemical preservatives have led to a special focus on the use of natural compounds, especially plant derivatives. The aim of this study was to investigate the antifungal properties of herbal essential oils of Cinnamon, Clove, Thymes, and Zataria multiflora, Cumin, and Caraway on the growth of Aspergillus ochraceus. Materials and Methods: Briefly, the presence of mycotoxin producing gene was investigated using PCR. Furthermore, production of mycotoxin in a medium with high performance liquid chromatography with fluorescence detection (FLD) was evaluated. Ingredients of essential oils were determined using GC/MS. The amount of antifungal activity of essential oils was assessed by disc diffusion and well diffusion method. Additionally, the minimum inhibitory concentration (MIC) was determined by macrodilution method. Results: Gene presence and mycotoxin production were confirmed by PCR and HPLC-FL. Among all studied essential oils, Cinnamon with MIC and MFC of 0.078 μl / ml exhibited the greatest effect on A. ochraceus growth as compared to other essential oils. Conclusion: This study indicated that essential oils have an effective role on controlling A. ochraceus growth and have shown promising to be a good bioactive natural preservative in food industry.

This study investigated the influence of temperature and water activity (aw) on the growth and biosynthesis of Ochratoxin A (OTA) by Aspergillus carbonarius and Aspergillus ochraceus, and the influence of climate change on the production potential of this mycotoxin in coffee was predicted. Central Composite Rotatable Design with aw ranging from 0.91 to 0.99 and temperatures from 17 to 38 °C was used. A. carbonarius CCDCA 10288 and 10293 showed optimal growth in aw intervals from 0.935 to 0.965 and temperatures between 25 and 32 °C while A. ochraceus CCDCA 10211 and 10212 showed aw intervals from 0.940 to 0.990 and temperatures between 21 and 30 °C. The conditions under which strains show the highest OTA risk index are as follows: for A. carbonarius CCDCA 10288 and 10293, aw between 0.95 and 0.99 and temperatures between 22 and 32 °C, for A. ochraceus CCDCA 10211 aw between 0.97 and 0.99 and temperatures between 25 and 30 °C. Using the models obtained, it was possible to verify that the conditions become more favorable for OTA production in the harvest months due to increased temperatures in the southern region of Minas Gerais. A safe product can be ensured by following good practices.

AIMS As a natural antimicrobial agent, Melaleuca alternifolia oil (MAO) is generally recognized to be safe and effective in the inhibition of phytopathogenic fungi. Due to lack of comprehensive studies on MAO for controlling postharvest Aspergillus, we investigated the preservative mechanism of MAO and its components against Aspergillus ochraceus in postharvest grapes to evaluate their potential effectiveness as fruit preservatives. METHODS AND RESULTS In our study, the compositions in MAO were analysed by gas chromatography-mass spectrometry. The inhibitory effects of MAO and its main constituents against A. ochraceus were compared by scanning electron microscopy and transmission electron microscopy observation, and metabolic analysis. Two components of MAO, α-terpineol and terpene-4-alcohol, showed higher antifungal effects than MAO, of which α-terpineol caused the worst leakage of cytoplasm and most serious hyphae distortions and spore disruptions. The downregulation of metabolic pathways of A. ochraceus was strongest with α-terpineol. The best inhibitory efficacy against A. ochraceus in grapes also occurred with α-terpineol. 3-Carene showed little inhibitory effect. CONCLUSIONS These results demonstrate that not all components in MAO possess antimicrobial effects, and α-terpineol is the main contributor of MAO's A. ochraceus inhibition effect. SIGNIFICANCE AND IMPACT OF THE STUDY α-Terpineol may be used as an alternative natural preservative for the postharvest storage of grapes and other fruits.

Aspergillus spp. are ubiquitous fungi that grow on stored grains. Some species produce toxins that can harm human and animal health, leading to hepato- and nephrotoxicity, immunosuppression and carcinogenicity. Major fungicides used to prevent fungal growth may be toxic to humans and their repeated use over time increases levels of resistance by microorganisms. Nanotechnology is an emerging field that allows use of antimicrobial compounds in a more efficient manner. In this study, was evaluated the antifungal activity of biogenic silver nanoparticles (AgNPs, synthesized by fungi) and simvastatin (SIM, a semi-synthetic drug), alone and in combination against three toxigenic species belonging to the genera Aspergillus section Flavi (Aspergillus flavus, Aspergillus nomius and Aspergillus. parasiticus) and two of section Circumdati (Aspergillus ochraceus and Aspergillus melleus). SIM exhibited a MIC50 of 78 μg/mL against species of Section Flavi and a MIC50 of 19.5 μg/mL against species of Section Circumdati. The MIC50 of AgNPs against Aspergillus flavus, Aspergillus nomius and Aspergillus parasiticus was 8 μg/mL, while the MIC50 was 4 μg/mL against Aspergillus melleus and Aspergillus ochraceus. Checkerboard assay showed that these compounds, used alone and in combination, have synergistic and additive effects against toxicogenic species of Aspergillus. Analysis by SEM gives an idea of the effect of SIM and AgNPs alone and in combination on spore germination and vegetative growth. Ultrastructural analysis revealed that spore germination was prevented, or aberrant hyphae were formed with multilateral branches upon treatment with SIM and AgNPs. These results reveal potential benefits of using combination of AgNPs and SIM to control fungal growth.

Symbiotic plant-fungi interaction is a promising approach to alleviate salt stress in plants. Moreover, endophytic fungi are well known to promote the growth of various crop plants. Herein, seven fungal endophytes were screened for salt tolerance; the results revealed that Aspergillus ochraceus showed a great potentiality in terms of salt tolerance, up to 200 g L−1. The indole acetic acid (IAA) production antioxidant capacity and antifungal activity of A. ochraceus were evaluated, in vitro, under two levels of seawater stress, 15 and 30% (v/v; seawater/distilled water). The results illustrated that A. ochraceus could produce about 146 and 176 µg mL−1 IAA in 15 and 30% seawater, respectively. The yield of IAA by A. ochraceus at 30% seawater was significantly higher at all tryptophan concentrations, as compared with that at 15% seawater. Moreover, the antioxidant activity of ethyl acetate extract of A. ochraceus (1000 µg mL−1) at 15 and 30% seawater was 95.83 ± 1.25 and 98.33 ± 0.57%, respectively. Crude extracts of A. ochraceus obtained at 15 and 30% seawater exhibited significant antifungal activity against F. oxysporum, compared to distilled water. The irrigation of barley plants with seawater (15 and 30%) caused notable declines in most morphological indices, pigments, sugars, proteins, and yield characteristics, while increasing the contents of proline, malondialdehyde, and hydrogen peroxide and the activities of antioxidant enzymes. On the other hand, the application of A. ochraceus mitigated the harmful effects of seawater on the growth and physiology of barley plants. Therefore, this study suggests that the endophytic fungus A. ochraceus MT089958 could be applied as a strategy for mitigating the stress imposed by seawater irrigation in barley plants and, therefore, improving crop growth and productivity.

Mycoremediation is an important process that targets the removal of petroleum hydrocarbons by fungi. Accordingly, colorimetric method was used in the preliminary investigation of petroleum degradation with ten fungal strains as Aspergillus ochraceus, Aspergillus parasiticus, Aspergillus niger, Fusarium acuminatum, Fusarium graminearum, Fusarium equiseti, Fusarium oxysporum, Paecilomyces lilac, Penicillium crustosum, and Penicillium chrysogenum. Petroleum degradation of spore suspension, live biomass (fungal pellet and disc) and cell-free culture supernatant of the potent A. ochraceus strain were determined by gravimetric analysis. It was obtained that the fungal disc (94%) was more successful than the spore suspension (87%) in petroleum degradation under optimized conditions as pH:5.0, 1% of petroleum concentration, %5 (v/v) of inocum concentration, 1 g/100mL of inoculum amount and 7 days of incubation period. The degradation rate constant and half-life period of spore suspension were calculated as 0.291 day− 1 and t1/2 = 0.340 and of fungal disc were 0.401 day− 1 and t1/2 = 0.247. 7.5% and 10% (v/v) concentration of cell-free culture supernatant were achieved more than 80% of petroleum removal. However, the cell-free culture supernatant was not as effective as fungal disc. According to GC/MS analysis, the fungal disc of A. ochraceus strain degraded long chain n-alkanes such as C35 and C36 more effectively than n-alkanes in the range of C22-C34. Drop-collapse and oil-spreading methods showed that A. ochraceus is a good biosurfactant producer. This study clearly pointed out that Aspergillus ochraceus NRRL 3174 strain with high its removal capacity can be used as an effective agent in petroleum bioremediation process.

The biological activity of essential oils and their major components is well documented. Essential oils such as oregano and cinnamon are known for their effect against bacteria, fungi, and even viruses. The mechanism of action is proposed to be related to membrane and external cell structures, including cell walls. This study aimed to evaluate the biological activity of seven essential oils and eight of their major components against Gram-negative and Gram-positive bacteria, filamentous fungi, and protozoans. The antimicrobial activity was evaluated by determination of the Minimal Inhibitory Concentration for Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella Typhimurium, Shigella sonnei, Aspergillus niger, Aspergillus ochraceus, Alternaria alternata, and Fusarium oxysporium, the half-maximal inhibitory concentration (IC50) for Trypanosoma cruzi and Leishmania mexicana, and the median lethal dose (LD50) for Giardia lamblia. Results showed that oregano essential oil showed the best antibacterial activity (66–100 µg/mL), while cinnamon essential oil had the best fungicidal activity (66–116 µg/mL), and both showed excellent antiprotozoal activity (22–108 µg/mL). Regarding the major components, thymol and carvacrol were also good antimicrobials (23–200 µg/mL), and cinnamaldehyde was an antifungal compound (41–75 µg/mL). The major components were grouped according to their chemical structure as phenylpropanoids, terpenoids, and terpinenes. The statistical analysis of the grouped data demonstrated that protozoans were more susceptible to the essential oils, followed by fungi, Gram-positive bacteria, and Gram-negative bacteria. The analysis for the major components showed that the most resistant microbial group was fungi, which was followed by bacteria, and protozoans were also more susceptible. Principal Component Analysis for the essential oils demonstrated the relationship between the biological activity and the microbial group tested, with the first three components explaining 94.3% of the data variability. The chemical structure of the major components was also related to the biological activity presented against the microbial groups tested, where the three first principal components accounted for 91.9% of the variability. The external structures and the characteristics of the cell membranes in the different microbial groups are determinant for their susceptibility to essential oils and their major components

The article reports the chemical composition, antioxidant, six key enzymes inhibitory and antimicrobial activities of two solvent extracts (water and methanol) of leaves and stem bark of Uapaca togoensis. For chemical composition, methanol extract of stem bark exhibited significant higher total phenolic (129.86 mg GAE/g) and flavanol (10.44 mg CE/g) contents. Methanol extract of leaves and water extract of stem bark showed high flavonoids (20.94 mg RE/g) and phenolic acid (90.40 mg CAE/g) content, respectively. In addition, HPLC-ESI-TOF-MS analysis revealed that U. togoensis was rich in procyanidins. The methanol and water extracts of stem bark had overall superior antioxidant activity; however, only methanol extract of stem bark showed higher inhibition of cholinesterase (AChE: 2.57 mg GALAE/g; BChE: 4.69 mg GALAE/g), tyrosinase (69.53 mg KAE/g) and elastase (2.73 mmol CE/g). Potent metal chelating ability was showed by water extract of leaves (18.94 mg EDTAE/g), higher inhibition of amylase was detected for water extracts of leaves (0.94 mmol ACAE/g) and stem bark (0.92 mmol ACAE/g). The tested extracts have shown wide-spectrum antibacterial properties and these effects have shown to be more effective against Aspergillus ochraceus, Penicillium funiculosum, Trichoderma viride, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. The results revealed that the antioxidant, enzyme inhibitory and antimicrobial activities depended on the extraction solvents and the parts of plant. Bioinformatics analysis on the 17 major compounds showed modulation of pathway associated with cancer. In brief, U. togoensis might be valuable as potential source of natural agents for therapeutic application.

BackgroundMeat-products are considered an enriched media for mycotoxins. This study aimed to investigate the prevalence of toxigenicAspergillusspecies in processed meat samples, HPLC-quantitative measurement of aflatoxin B1and ochratoxin A residues, and molecular sequencing ofaflR1andpksgenes. One hundred and twenty processed beef meat specimens (basterma, sausage, and minced meat;n = 40 for each) were collected from Ismailia Province, Egypt. Samples were prepared for total mold count, isolation, and identification ofAspergillusspecies. All samples were analyzed for the production of both Aflatoxin B1and Ochratoxin A mycotoxins by HPLC. Molecular identification ofAspergillus flavusandAspergillus ochraceuswas performed using PCR amplification of the internal transcribed spacer (ITS) region; furthermore, theaflR1andpksgenes were sequenced.ResultsThe total mold count obtained from sausage samples was the highest one, followed by minced meat samples. The prevalence ofA. flavuswas (15%), (7.5%), and (10%), while the prevalence ofA. ochraceuswas (2.5%), (10%), and (0%) in the examined basterma, sausage, and minced meat samples, respectively. Using PCR, the ITS region was successfully amplified in all the testedA. flavusandA. ochraceusstrains. Aflatoxin B1was detected in six basterma samples (15%). Moreover, the ochratoxin A was detected only in four sausage samples (10%). TheaflR1andpksgenes were amplified and sequenced successfully and deposited in the GenBank with accession numbers MF694264 and MF694264, respectively.ConclusionsTo the best of our knowledge, this is the first report concerning the HPLC-Molecular-based approaches for the detection of aflatoxin B1and ochratoxin A in processed beef meat in Egypt. The production of aflatoxin B1and ochratoxin A in processed meat constitutes a public health threat. Aflatoxin B1is commonly associated with basterma samples. Moreover, ochratoxin A was detected frequently in sausage samples. The routine inspection of mycotoxins in processed meat products is essential to protect human consumers.

The purpose of this study was to evaluate the inhibitory effect of allyl-isothiocyanate (AITC) and benzyl-isothiocyanate (BITC) on fungal growth and Ochratoxin A (OTA) production by Aspergillus ochraceus, A. carbonarius and A. niger. Here, we found that spore germination and fungal growth of the three fungi were significantly inhibited when the concentration of AITC and BITC was higher than 1.25 μg/mL. The inhibitory effect of AITC or BITC on A. carbonaceus and A. ochraceus was significantly stronger than that of A. niger. Scanning electron microscopy showed that the mycelia of all three fungi were changed by AITC and BITC. Compared with A. ochraceus and A. carbonarius, the damage to A. niger was lower. For OTA production, AITC and BITC could significantly down-regulated the expression of all five OTA biosynthesis genes in A. niger and A. carbonarius. In A. ochraceus, although several OTA biosynthesis genes were up-regulated, the key PKS gene was down-regulated by AITC and BITC. Twenty-five μg/mL of AITC or BITC could reduce the infection of the three fungi on grapes with inhibition rates of 28%–36% during 14 days and prolong the shelf life of grapes. In maize, the OTA production of the three fungi was significantly reduced by 25 μg/mL of AITC and BITC with the inhibition rates 68.04%–93.49% and 65.87%–75.45%, respectively. These results suggest that AITC and BITC can be used as natural fungicides to prevent A. niger, A. carbonarius and A. ochraceus from infecting grapes and maize and control OTA contamination.

This study was carried out to evaluate the prevalence of mycobiota and aflatoxin (AF) B1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. For this purpose, 100 sesame seed samples were collected directly from the fields of major sesame-producing areas in rainfed and irrigated zones. The agar plate method was used for isolation of mycobiota, and thin-layer chromatography was used to determine AFB1 concentrations. Seed samples were then stored for 12 months. After 12 months, the seeds were again analyzed for mycobiota and AFB1 for comparison. All samples were positive for fungal growth under fresh and stored conditions. Twenty-one fungal species of 10 genera were isolated. Aspergillus flavus was the most prevalent contaminant found in fresh and stored sesame seeds from rainfed and irrigated zones, followed by Aspergillus niger, Alternaria alternata, and Fusarium oxysporum. The least prevalent fungi were Aspergillus ochraceus and Cladosporium oxysporum. Analysis revealed that 92% of fresh and 99% of stored seed samples were contaminated with AFB1. In the rainfed zone, 88% of fresh and 100% of stored seed samples were contaminated with AFB1, with mean concentrations of 15.74 and 33.8 ppb, respectively. In the irrigated zone, 96% of fresh and 98% of stored seed samples were contaminated with AFB1, with mean concentrations of 20.5 and 27.56 ppb, respectively. AFB1 concentrations >20 ppb were found in 20% of fresh and 100% of stored seeds samples from the rainfed zone and in 28% of fresh and 60% of stored samples from the irrigated zone and thus were not fit for human consumption as per the maximum limit (20 ppb) assigned by the U.S. Food and Drug Administration and the Food and Agriculture Organization of the United Nations. This report is the first on the mycobiota and AFB1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue. HIGHLIGHTS

Grains are susceptible to contamination by molds; some cause spoilage and others produce certain mycotoxins that cause a serious health threat to humans and animals. Aspergillus flavus and Aspergillus ochraceus and their mycotoxins, aflatoxins and ochratoxin A, are natural contaminants of various agricultural commodities. Control of these molds and their mycotoxins in food commodities is of utmost importance; therefore, the target of this research was to explore the effects of gamma irradiation doses on the growth of A. flavus and A. ochraceus in artificially inoculated yellow maize as well as on the production of aflatoxin B1, ochratoxin A, and the formation of toxins in maize. The irradiated dose of 6.0 kGy was found to completely inhibit the growth of the two molds, while a dose of 4.5 kGy reduced the production of their mycotoxins. Maximum degradation of the formed aflatoxins and ochratoxin A in maize occurred at 20 kGy, with best reduction rates of 40.1%, 33.3%, and 61.1% observed for aflatoxin B1, aflatoxin B2, and ochratoxin A, respectively. We recommend grains irradiation by gamma radiation at 6.0 kGy to decontaminate mycotoxin-producing molds before they produce mycotoxins. The study represents a proactive, efficient, and potent method for avoiding potential contamination of fungus during grains storage and transfer for one to two months.

Fungal infection and synthesis of mycotoxins in coffee leads to significant economic losses. This study aimed to investigate the prevalence of toxigenic fungi, their metabolites, and the effect of traditional roasting and brewing on ochratoxin A (OTA) and aflatoxins (AFs) contents of naturally contaminated coffee samples. In addition, in vivo biocontrol assays were performed to explore the antagonistic activities of Bacillus simplex 350–3 (BS350-3) on the growth and mycotoxins synthesis of Aspergillus ochraceus and A. flavus. The relative density of A. niger, A. flavus, Penicillium verrucosum and A. carbonarius on green coffee bean was 60.82%, 7.21%, 3.09% and 1.03%, respectively. OTA contents were lowest in green coffee beans (2.15 μg/kg), followed by roasted (2.76 μg/kg) and soluble coffee (8.95 μg/kg). Likewise, AFs levels were highest in soluble coffee (90.58 μg/kg) followed by roasted (33.61 μg/kg) and green coffee (9.07 μg/kg). Roasting naturally contaminated coffee beans at three traditional methods; low, medium and high, followed by brewing resulted in reduction of 58.74% (3.50 μg/kg), 60.88% (3.72 μg/kg) and 64.70% (4.11 μg/kg) in OTA and 40.18% (34.65 μg/kg), 47.86% (41.17 μg/kg) and 62.38% (53.73 μg/kg) AFs contents, respectively. Significant inhibitions of AFs and OTA synthesis by A. flavus and A. carbonarius, respectively, on infected coffee beans were observed in presence of Bacillus simplex BS350-3 volatiles. Gas chromatography mass spectrochemistry (GC-MS/MS) analysis of head-space BS350-3 volatiles showed quinoline, benzenemethanamine and 1-Octadecene as bioactive antifungal molecules. These findings suggest that marketed coffee samples are generally contaminated with OTA and AFs, with a significant level of roasted and soluble coffee contaminated above EU permissible limits for OTA. Further, along with coffee roasting and brewing; microbial volatiles can be optimized to minimize the dietary exposure to mycotoxins.

Background Aspergillus ochraceus causes food spoilage and produces mycotoxin ochratoxin A (OTA) during storage of agricultural commodities. In this study, citral was used to inhibit A. ochraceus growth and OTA accumulation, proteomic analysis was employed to verify the mechanism of citral. Results Citral was found to significantly inhibit fungal growth and mycotoxin production in A. ochraceus. Specifically, 75, 125, 150 and 200 μL L-1 citral suppressed mycelial growth by 33%, 46%, 50% and 100%, respectively. Additionally, 75 μL L-1 citral inhibited OTA accumulation by 25%. Proteomic analysis was performed to elucidate the inhibitory mechanism of citral on mycelial growth and OTA production at subinhibitory concentrations (75 μL L-1 ). Proteomics analysis identified 2646 proteins in A. ochraceus fc-1, of which 218 were differentially expressed between control and 75 μL L-1 citral treatment samples. Differentially expressed proteins were identified by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of biological process, cellular component and molecular function terms. Potential factors affecting mycelial growth and OTA production were analysed, and OTA production was revealed to be a complex process involving many associated factors related to various processes including nutrient intake, sterol biosynthesis, ribosome biogenesis, energy metabolism, oxidative stress and amino acid metabolism. In addition, citral at 75 μL L-1 down-regulated OTA biosynthetic genes including pks and nrps, but slightly up-regulated the global regulatory factors veA, velB and laeA. Conclusion The findings further demonstrate the potential of citral for the preservation of grains and other agricultural products, and provide new insight into its antifungal mechanisms at subinhibitory concentrations. © 2021 Society of Chemical Industry.

Antifungal and antibacterial activities of crude extracts of 3 tropical mushrooms including Pleurotus sajor-caju, Pleurotus tuber-regium and Lentinus squarrosulus were investigated on eleven species of bacterial and three of fungal human pathogens. For the pathogenic fungi, the Minimal Inhibitory Concentration (MIC) of carpophore extracts ranged from 0.39 mg/mL to 6.25 mg/mL for Candida albicans, 0.78 mg/mL to 6.25 mg/mL for Aspergillus fumigetus, and 1.56 mg/mL to 6.25 mg/mL for Aspergillus ochraceus. For bacteria, the MIC values ranged from 6.25 mg/mL to 12.5 mg/mL on most Gram positive strains including Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus and Mycobacterium smegmatis. This MIC value was the same (12.5 mg/mL) for the 3 crude extracts tested on Staphylococcus epidermidis for the Gram positive strains. Gram negative bacteria were generally less sensitive to crude extracts with higher MIC values ranging from 6.25 mg/mL to 12.5 mg/mL for Escherichia coli and Enterobacter cloacae and the same (12.5 mg/mL) for Proteus vulgaris, Klebsiella oxytoca, Klebsiella aerogenes and Proteus mirabilis. Based on the above mentioned figures, it appears that strains of pathogenic fungi tested are generally much more sensitive to crude extracts than strains of bacteria. In fact, antimicrobial activities of the 3 crude extracts tested are stronger on human pathogenic fungi than bacteria. These results are evidence that carpophores of the 3 mushrooms species could be a source of new molecules potentially more effective than synthetic products against some human pathogenic fungi and bacteria.

The study was conducted to evaluate the activity of acetic acid and sodium bicarbonate to inhibit Aspergillus ochraceus on culture media and reduce ochra toxin A in poultry diet. It was found that addition of acetic acid into PDA medium 0.2%, 0.5% and 4% could reduce A ochraceus growth by 90.22%, 98.77and 100%, respectively. Similar reduction in A ochraceus growth was obtained on PDA containing sodium bicarbonate that attained to 81%, 91.5%, and 100% at 4%, 2% and 5% concentrations, respectively. Results showed that addition of acetic acid at 0.5% and sodium bicarbonate at 1% could reduce ochre A mycotoxin by 65.7% and 43.6%, respectively in the first month and then reached to 100% in the second month for the two compounds.

Highlights • Vermiculite is a new promising carrier for solid-state fermentation of micromycetes. • Aspergillus have higher protease production on vermiculite in comparison with on other carriers. • The mycelium productivity of Aspergillus was higher not only in comparison with SmF, but also in comparison with other carriers under SSF conditions (from 3 to 18 times)., A new method has been developed to increase the productivity of aspergilli - producers of extracellular proteinases based on their cultivation on vermiculite under solid-state fermentation conditions. The productivity of the mycelium Aspergillus ochraceus L-1 and Aspergillus ustus 1 was 3-18 times higher not only in comparison with submerged cultivation, but also in comparison with growth on other carriers studied under solid-state fermentation conditions. Vermiculite can be considered as a new promising carrier for solid-state fermentation of micromycetes.

Aim is to study the effects of aqueous and methanolic of fruits leaves and roots of Citrullus colocynths Schard on fungal strain. Two methods have been used with this extracts on 2 fungal strains producer of mycotoxins; determination of growth radial technique on solid medium and determination of biomass technique on liquid medium. Aqueous and methanolic extract of fruit, leaves and root of Citrullus colocynths Schard; these are examined for their antifungal potentiality against Aspergillus flavus and Aspergillus ochraceous.

Hurdle technologies are among the novel food processing techniques for food preservation, where there is a combination of mild antimicrobial factors, which can interact on the inhibition of microbial spoilage. Spices essential oils are widely known as antimicrobial compounds that can have a preservation effect on food, but their effect can be affected by food components. In order to evaluate the effect of pH (4.5-7.0) and protein content (2-8%) on the antifungal effect of Mexican oregano essential oil (150-600 mg/L), a Box-Benkhen design was used to test the effect of the treatments on the radial growth of two food-related fungi, Penicillium expansum (NRRL 2304) and Aspergillus ochraceus (ATCC 22947). Three essential oils were used, these with a different chemical composition. The first chemotype of Lippia berlandieri Schauer, presented a higher proportion of carvacrol (58.23%). The second Lippia berlandieri Schauer essential oil presented a higher concentration of thymol (60.15%). Essential oil from Poliomintha longiflora Gray contained a 43.02% of thymol. Kinetic growth parameters were obtained using the modified Gompertz equation. The maximum mold growth (A) of the molds was affected in a lineal concentration-dependent effect by the three Mexican oregano essential oils and protein concentration. This phenomenon is independent of the chemistry variability in the composition of the three OEO. The higher concentration of protein (8%) was also related to an increase in lag time (λ). These two growth parameters were mainly statistically significant of all treatments evaluated, when considering as intrinsic parameters in food such as pH and protein content. The action of OEO was higher against A. ochraceus, positioning itself as the most sensitive microorganism. The use of predictive modelling can provide valuable information on the interaction of food components to enhance or decrease the antifungal capacity of essential oils.

Polyphenols, obtained from natural resources, may possess important pharmacological effects. The polyphenolic profiles of the stem extracts of six Ferocactus species (sp.): F. gracilis, F. pottsii, F. herrerae, F. horridus, F. glaucescens, and F. emoryi, were measured using high-performance liquid chromatography (HPLC) with diode-array detection (DAD). Additionally, anticancer, antibacterial, and antifungal activities were examined. Results showed the presence of high to moderate amounts of polyphenols in the extracts (phenolic acids: Protocatechuic acid, 3,4-dihydroxyphenylacetic acid, caffeic acid, and vanillic acid, flavonoids: Rutoside and quercitrin). The highest amounts of 3,4-dihydroxyphenylacetic acid were found in F. glaucescens ((132.09 mg 100 g&minus, 1 dry weight (DW)), F. pottsii (75.71 mg 100 g&minus, 1 DW), and F. emoryi (69.14 mg 100 g&minus, 1 DW) while rutoside content was highest in F. glaucescens (107.66 mg 100 g&minus, 1 DW). Maximum antiproliferative activities were observed against HeLa and Jurkat cancer cells, with F. glaucescens, F. emoryi, and F. pottsii showing the highest anticancer activity. Most bacteria were sensitive to Ferocactus sp. stem extracts. Escherichia coli and Staphylococcus aureus were the most sensitive. Excellent antifungal effects were observed against Aspergillus ochraceus and A. niger. However, Penicillium funiculosum, P. ochrochloron, and Candida albicans were relatively resistant. This is the first study reporting novel sources of polyphenols in Ferocactus sp. with anticancer and antimicrobial activities.

Recently, specific dry-cured hams have started to be produced in San Daniele and Parma areas. The ingredients are similar to protected denomination of origin (PDO) produced in San Daniele or Parma areas, and include pork leg, coming from pigs bred in the Italian peninsula, salt and spices. However, these specific new products cannot be marked as a PDO, either San Daniele or Parma dry cured ham, because they are seasoned for 6 months, and the mark PDO is given only to products seasoned over 13 months. Consequently, these products are called short-seasoned dry-cured ham (SSDCH) and are not branded PDO. During their seasoning period, particularly from the first drying until the end of the seasoning period, many molds, including Eurotium spp. and Penicillium spp., can grow on the surface and work together with other molds and tissue enzymes to produce a unique aroma. Both of these strains typically predominate over other molds. However, molds producing ochratoxins, such as Aspergillus ochraceus and Penicillium nordicum, can simultaneously grow and produce ochratoxin A (OTA). Consequently, these dry-cured hams may represent a potential health risk for consumers. Recently, Aspergillus westerdijkiae has been isolated from SSDCHs, which could represent a potential problem for consumers. Therefore, the aim of this study was to inhibit A. westerdijkiae using Debaryomyces hansenii or Lactobacillus buchneri or a mix of both microorganisms. Six D. hansenii and six L. buchneri strains were tested in vitro for their ability to inhibit A. westerdijkiae. The strains D. hansenii (DIAL)1 and L. buchneri (Lb)4 demonstrated the highest inhibitory activity and were selected for in situ tests. The strains were inoculated or co-inoculated on fresh pork legs for SSDCH production with OTA-producing A. westerdijkiae prior to the first drying and seasoning. At the end of seasoning (six months), OTA was not detected in the SSDCH treated with both microorganisms and their combination. Because both strains did not adversely affect the SSDCH odor or flavor, the combination of these strains are proposed for use as starters to inhibit OTA-producing A. westerdijkiae.

Aflatoxin M1 (AFM₁) and ochratoxin A (OTA) are two main mycotoxins in milk and dairy products. In the present work, the ability of four Lactobacillus strains (L. plantarum PTCC 1058, L. plantarum LP3, L. plantarum AF1 and L. plantarum LU5) to remove AFM1 and OTA in fermented cream was studied during 24 h fermentation. The antifungal activity of the mentioned lactobacilli against the defined fungi (Aspergillus flavus PTCC 5004, Aspergillus parasiticus PTCC 5018, Aspergillus nidulans PTCC 5014, Aspergillus ochraceus PTCC 5060) was also evaluated. The results showed that the cell counts of all strains were increased by 64–70% during fermentation. All Lactobacillus strains decreased the amount of AFM₁ significantly (P ≤ 0.05) in the range of 26–52%, which the highest AFM₁‐reducing effect was related to L. plantarum LU5 (from 0.5 to 0.24 μg kg⁻¹). The mean OTA removal by Lactobacillus strains in fermented cream also ranged from 32 to 58%. Amongst Lactobacillus strains, the cell‐free culture supernatants of L. plantarum LU5 showed the highest (inhibition zone of 26.7 ± 1.2 mm) and L. plantarum LP3 and L. plantarum PTCC 1058 the lowest antifungal activities. The fermented creams contained Lactobacillus strains exhibited the highest and lowest antifungal activities against A. ochraceus and A. parasiticus, respectively. L. plantarum LU5, with the inhibition zone of 27.6 ± 0.9 mm, was the most effective fungal inhibitor, while L. plantarum PTCC 1058 had the lowest antifungal activity.

The development of effective control systems for post-harvest preservation and storage processes seems to be one of the best ways to reduce the activity of fungi and thus the risk of mycotoxin contamination of grain during its storage. A crucial step towards developing such a system requires the understanding of ecological relationships between environmental factors, fungal development and mycotoxin production in bulk of grain treated as holistic systems. The aim of this study was to estimate the critical points associated with mould development and mycotoxin production in the ecosystem of malting barley grain with a hazardous initial microbiological state (provided by the inoculation of naturally contaminated grain with Aspergillus ochraceus and Penicillium verrucosum) stored in different temperature and water activity conditions (t = 12–30 °C and aw = 0.78–0.96, where aw corresponds to equilibrium relative air humidity in inter-grain spaces). The most intensive fungal growth occurred in the ecosystems of barley grain with aw = 0.91–0.93 stored at 24 and 30 °C. The biosynthesis of ochratoxin A (OTA) was observed at 0.91–0.95 aw at 18–30 °C, whilst penicillic acid and citrinin were detected only at temperatures of 24 and 30 °C. The toxicogenic ability of fungi at 12–24 °C and aw≤0.80 was significantly limited. In the ecosystem of barley grain, the threshold of fungal concentration (expressed in colony forming units of moulds, CFU g−1), for which the amount of OTA exceeded binding limits, was 105 CFU g−1 of grain. The study indicates that the duration of the lag phase of fungal development (comprising spores germination and the beginning of exponential mould growth), which in all tested ecosystems was shorter than the time, after which admissible levels of OTA were exceeded, may be used to evaluate the safe period, at which grain should be subjected to preservation processes.

A strain of Aspergillus ochraceus was isolated from the stored low denatured soybean meals (LDSMs) and an extracellular protease was purified from the fungus grown culture to electrophoretical homogeneity. The protease had a molecular weight of approximately 70 kDa and showed proteolytic activities only in the presence of activators. The maximum activity was obtained in 1.4% sodium dodecyl sulfate (SDS) solution but similar activity could also be achieved in the presence of linoleic acid at the concentration as low as 0.05%. The protease was stable at pH 9.0–12.0 and remained active in the presence of methanol, ethanol, acetone and trichloromethane and was classified as serine protease. The thermal stability of the protease located in the temperature range of 4–40 °C and an activator independent autolysis was observed above 60 °C. Soy proteins was hydrolyzed by the purified protease in a model system using residual lipids from stored LDSMs as the activator and a degree of hydrolysis (DH) of 6.01% ± 0.27 was obtained after reaction for 60 min at pH 9.0 and 40 °C. This study gives new insights into the quality control of the soy protein production and the use of fungal proteases in the modification of soy proteins.

Proteases have different industrial applications thus to increase their yield and reduce the production cost; a low cost fermentation medium was designed for large scale production. From seven tested agro-wastes, wheat bran (1.0 g%) was found to increase the overall yield of alkaline protease from Aspergillus ochraceus BT21 more than 2 fold (7359.8 U/mg) compared to the control (optimized fermentation conditions obtained from our previous study). The enzyme was purified to homogeneity using ammonium sulphate precipitation, Sephacryl S-200, followed by DEAE-Sepharose and CM-Sepharose chromatography. The efficiency of the purification steps was confirmed by SDS–PAGE where, the molecular mass of the enzyme was 59.0 KDa. The purification fold was 15.13 with specific activity reached 111379.5 U/mg and 59.7% recovery. The enzyme was alkaline thermo-tolerant with optimum values of temperature and pH at 50 °C and 8.0, respectively. The purified enzyme was stable in a wide range of temperature (20–60 °C) and pH (5.0–11.0), and its maximum activity was observed with casein substrate. The inhibition studies showed that it is a thiol-dependant serine protease. The enzyme exhibited good stability with the commercial detergents. The specific properties of the purified enzyme suggested it is suitable for future industrial applications especially in detergent formulations.

Temperature and water activity (aw) are both responsible for the growth and production of ochratoxin A (OTA) by Aspergillus species in coffee. To prove this, we tested the effect of environmental factors combined with green coffee composition on the risk of OTA production as well as the efficiency of using Saccharomyces cerevisiae to control the growth of Aspergillus ochraceus, A. westerdijkiae, and A. carbonarius, OTA production and biosynthetic genes expression regulation levels. The results showed a higher growth rate and OTA risk index (ORI) for the three fungal isolates at 21 °C and 0.98 aw. Temperature affected the growth of the isolates negatively. The ORI was influenced synergistically by aw and antagonistically by temperature. The best condition for biological control by using S. cerevisiae CCMA 0159 was observed at 28 °C and 0.99 aw. Temperature and aw directly affected OTA production and biological control efficiency, but at different values. A. carbonarius CCDCA 10507 showed better growth and a greater risk of contamination in coffee. S. cerevisiae CCMA 0159 significantly reduced the risk of contamination by Aspergillus using substrate competition as strategies, acting on down regulation of OTA's biosynthetic gene expression, including hal and p450.

The aim of this work was the study of the main effects of benzyl isothiocyanate (BITC) on A. ochraceus morphology and on its production metabolism of ochratoxins. This compound was evaluated as active agent of an antimicrobial label in food packaging. Microbiological studies showed a slowdown in mould growth when the active material was applied to A. ochraceus and the presence of three different areas of growth. Scanning electron microscopy was successfully used to demonstrate the mode of action of BITC on this strain. A. ochraceus exhibited modifications in morphology compared to the control samples such as the disappearance of sclerotia or cleistothecia An extraction protocol and an analytical method by UPLC-MS/MS to determine ochratoxins (OTs) was developed. The results showed that all these morphological changes were related to a decrease on OTs production, both ochratoxin A (OTA) and ochratoxin B (OTB). The presence of BITC caused a great decrease on OTA that modified the OTA/OTB ratio, increasing the OTB proportion. Furthermore, the active packaging also modified the production of other secondary metabolites. The morphological and metabolic effects observed, as well as the relationship between them, are of great interest since they have not been reported before for A. ochraceus.

One of the emerging concerns in the Cordillera Administrative Region, Philippines is ochratoxin A (OTA) contamination in coffee. During 2015 to 2016, a total of 51 Arabica (Coffea arabica) coffee samples from Benguet province and 71 Robusta (Coffea canephora var. Robusta) coffee samples from the provinces of Ifugao and Kalinga were analysed for OTA contamination. The OTA-producing fungal contaminants during drying and storage of Arabica and Robusta coffee were Aspergillus niger and Aspergillus ochraceus. Ochratoxin A was more commonly detected in Robusta coffee (36.6%) than in Arabica coffee (21.6%). Among the contaminated samples, Robusta coffee cherries in the drying yard had the highest mean OTA level (120.2 μg kg-1, n = 10) while roasted Robusta coffee beans had the lowest mean level (4.8 μg kg-1, n = 9). The onset of contamination of Arabica coffee occurred during storage, with a mean OTA level of 46.7 μg kg-1 (n = 9). Roasted coffee had lower OTA content although five samples had levels >5.0 μg kg-1. Pearson Chi-square analysis (χ2) and Fisher's exact test revealed that several post-harvest practices involving non-removal of the husk or hull and mixing of defective coffee were significantly associated with the occurrence of OTA during drying and storage (p