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Simple Summary: Ochratoxin A (OTA), a nephrotoxic mycotoxin categorized as a possible carcinogenic agent, is a current problem in the cheese manufacturing industry. The main OTA producers are Penicillium nordicum and Penicillium verrucosum. Therefore, we aimed to investigate different cheese ripening phases to hinder OTA production. We elaborated cheese analogues, inoculated both fungal species, and performed two different ripening phases, at 10 and 15 °C. Despite the fact that both were able to grow, they did not express genes related to OTA production, and no OTA was detected during two weeks of ripening. Therefore, we show that, when the first phases of ripening are maintained at <15 °C, the possibility of OTA production is dramatically minimized. This information is of utmost importance for industries to enhance the safety of their cheese products, which positively impacts human health. A lack of control of the technological abiotic parameters apparent during cheese manufacture, including temperature and relative humidity, results in this dairy product being prone to mold contamination. Sometimes, inoculant molds are used to obtain the characteristic sensory properties of this type of product. However, during the maturation process, some unwanted molds can colonize the ripening cheese and produce mycotoxins. Mycotoxigenic molds such as Penicillium nordicum and Penicillium verrucosum can colonize ripened cheeses, contaminating them with ochratoxin A (OTA), a nephrotoxic 2B toxin. Thus, the presence of OTA in cheeses could represent a hazard to consumers' health. This study has evaluated the growth and OTA production of P. nordicum and P. verrucosum on a cheese analogue under simulated ripening conditions of 10 and 15 °C and 0.96 water activity (aw). Ecophysiological, molecular, and analytical tools assessed the mold growth, gene expression, and OTA production under these environmental conditions. Both species were able to effectively colonize the cheese under these ripening conditions. However, neither species expressed the otapks and otanps biosynthetic genes or produced phenotypic OTA. Therefore, these results suggest a relatively low risk of exposure to OTA for consumers of this type of cheese product. The conditions used were thus appropriate for cheese ripening to minimize the potential for contamination with such mycotoxins. An appropriate adjustment of the technological ripening parameters during such cheese manufacture could contribute to OTA-free cheeses. [ABSTRACT FROM AUTHOR]

A lack of control of the technological abiotic parameters apparent during cheese manufacture, including temperature and relative humidity, results in this dairy product being prone to mold contamination. Sometimes, inoculant molds are used to obtain the characteristic sensory properties of this type of product. However, during the maturation process, some unwanted molds can colonize the ripening cheese and produce mycotoxins. Mycotoxigenic molds such as Penicillium nordicum and Penicillium verrucosum can colonize ripened cheeses, contaminating them with ochratoxin A (OTA), a nephrotoxic 2B toxin. Thus, the presence of OTA in cheeses could represent a hazard to consumers’ health. This study has evaluated the growth and OTA production of P. nordicum and P. verrucosum on a cheese analogue under simulated ripening conditions of 10 and 15 °C and 0.96 water activity (aw). Ecophysiological, molecular, and analytical tools assessed the mold growth, gene expression, and OTA production under these environmental conditions. Both species were able to effectively colonize the cheese under these ripening conditions. However, neither species expressed the otapks and otanps biosynthetic genes or produced phenotypic OTA. Therefore, these results suggest a relatively low risk of exposure to OTA for consumers of this type of cheese product. The conditions used were thus appropriate for cheese ripening to minimize the potential for contamination with such mycotoxins. An appropriate adjustment of the technological ripening parameters during such cheese manufacture could contribute to OTA-free cheeses.

The main goals of the present investigation were to develop O/W nanoemulsion fungicides based on cold-pressed Nigella sativa (black seed) oil to prevent Penicillium verrucosum infection of maize seeds and to test their antifungal activity against this fungus. Additionally, the effect of these nanoemulsions on plant physiological parameters was also investigated. Two nonionic surfactants namely Tween 20 and Tween 80 were used as emulsifying agents in these formulations. The effect of sonication time and surfactant type on the mean droplet size, polydispersity index (PDI), and zeta potential of the nanoemulsions were determined by dynamic light scattering (DLS). Results indicated that both sonication time and emulsifier type had pronounced effects on the stability of O/W nanoemulsions with a small particle size range (168.6–345.3 nm), acceptable PDI (0.181–0.353), and high zeta potential (−27.24 to –48.82 mV). Tween 20 showed superior stability compared to Tween 80 nanoemulsions. The in vitro results showed that complete inhibition of P. verrucosum-growth was obtained by 10_T80 and 10_T20 nanoemulsions at 100% concentration. All nanoemulsions had increment effects on maize seed germination by 101% in the case of 10_T20 and 10_T80 compared to untreated seeds or the chemical fungicide treatment. Nanoemulsions (10_T20 and 10_T80) were able to stimulate root and shoot length as compared to the fungicide treatment. Seed treatment with 10_T80 nanoemulsion showed the highest AI and protease activity by 75 and 70%, respectively, as compared to the infected control. The produced nanoemulsions might provide an effective protectant coating layer for the stored maize seeds. [ABSTRACT FROM AUTHOR]

The main goals of the present investigation were to develop O/W nanoemulsion fungicides based on cold-pressed Nigella sativa (black seed) oil to prevent Penicillium verrucosum infection of maize seeds and to test their antifungal activity against this fungus. Additionally, the effect of these nanoemulsions on plant physiological parameters was also investigated. Two nonionic surfactants namely Tween 20 and Tween 80 were used as emulsifying agents in these formulations. The effect of sonication time and surfactant type on the mean droplet size, polydispersity index (PDI), and zeta potential of the nanoemulsions were determined by dynamic light scattering (DLS). Results indicated that both sonication time and emulsifier type had pronounced effects on the stability of O/W nanoemulsions with a small particle size range (168.6–345.3 nm), acceptable PDI (0.181–0.353), and high zeta potential (−27.24 to –48.82 mV). Tween 20 showed superior stability compared to Tween 80 nanoemulsions. The in vitro results showed that complete inhibition of P. verrucosum-growth was obtained by 10_T80 and 10_T20 nanoemulsions at 100% concentration. All nanoemulsions had increment effects on maize seed germination by 101% in the case of 10_T20 and 10_T80 compared to untreated seeds or the chemical fungicide treatment. Nanoemulsions (10_T20 and 10_T80) were able to stimulate root and shoot length as compared to the fungicide treatment. Seed treatment with 10_T80 nanoemulsion showed the highest AI and protease activity by 75 and 70%, respectively, as compared to the infected control. The produced nanoemulsions might provide an effective protectant coating layer for the stored maize seeds.

The paper’s aim was to select the optimal mineral medium for LDPE biodegradation by the strain Penicillium verrucosum CNM-FP-02. It was selected five mineral salt media (MSM), which differed in salt content and N/P ratio. After 40 days of submerged cultivation, the following parameters were determined: catalase activity and pH of cultural media, biomass accumulation, rate of LDPE degradation, optical microscopy, and the tensile testing of polyethylene. It was observed that catalase activity on all media, except MSM 4, was higher in the presence of LDPE. The addition of polyethylene to the growth media stimulated the fungal biomass accumulation by 19.3-93.1% (4 media out of 5 tested). The percentage of degradation of LDPE films was different, depending on the culture medium, from 0.41% to 0.92%. The most active LDPE films were degraded on medium MSM 2. Visualization of plastic strips under an optical microscope revealed the immobilization of the microorganism and the damage on the polyethylene surface. The tensile test showed increased elasticity of the plastic in the variants treated with fungal strain. In conclusion, in order to stimulate the biodegradation of LDPE by the strain P. verrucosum CNM-FP-02, the medium MSM 2 (N/P ratio 1:1) was selected.

The current study aimed to detect fungi contamination in agricultural mushroom samples Agaricus bisporus which was taken from some areas of Baqubah district in Diyala governorate. It was shown that all the samples were contaminated 100%, total colony formation ranged 3.264×10-3 to 5.596 ×10-3, the results recorded 253 fungal isolates. The Canaan region recorded the highest number of fungal isolates, which was 84 isolates, followed by the regions of Buhruz, Khan Bani Saad and Baqubah, which were 66, 54 and 49 fungal isolates, in that order. The results of the study also showed that the fungal isolates that were isolated from the white agricultural fungus belonged to 3 fungal genera, called by Aspergillus, Rhizopus and Penicilium. The genus Aspergillus was superior in the number of species belonging to it since 6 species were recorded at a rate of 75% compared to the genus Rhizopus for which one species was recordedonly Rhizopus spp. The genus Penicillium recorded one species with a rate of 12.5 % each. The total species that were isolated and diagnosed were 8 fungal species: Aspergillus niger, A. parasiticus, A. nidulans, A. tamarii, A. fumigatus, A.ochraceus, Rizopus spp. and Penicillium verrucosum. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a nephrotoxin that contaminates grains in storage. Moisture and temperature sensors give delayed responses due to their slow kinetic movement within the silo. This study examines if CO 2 production could predict OTA contamination and identify storage conditions exceeding the maximum limit (5 μg/kg). The impact of water activity levels (0.70–0.90 a w), temperatures (15 and 20 °C), and storage duration on (a) Penicillium verrucosum population, (b)CO 2 respiration rates (RR), and (c)ochratoxins concentrations in stored wheat was investigated. 96 samples were analysed for ochratoxins with LCMS-MS. RR was >7 times higher at wetter conditions than at drier a w levels. A positive correlation between CO 2 , OTA, OTB, and OTα was observed at the wettest conditions. OTA exceeded the limit at >0.80 a w (16% moisture content) with RR > 0.01 mg CO 2 kg−1 h−1. The knowledge of the RR of stored grain would alert grain farmers/managers to improve grain storage management. • CO 2 respiration rates will increase as the water activity (a w) of grains increases. • a w influences Penicillium populations and OTA, OTB, and OTα concentrations in wheat. • CO 2 can be used to predict ochratoxin contamination in very wet grains. • Monitoring changes in CO 2 and abiotic factors will improve grain storage management. [ABSTRACT FROM AUTHOR]

The present study focused on phytofabrication of selenium nanoparticles (SeNPs) from Carica papaya extract and exploration of their multi-biofunctional features. Total phenolics and flavonoids of C. papaya fruit extract were determined as 23.30 ± 1.88 mg gallic acid equivalents and 19.21 ± 0.44 mg quercetin equivalents per gram, respectively, which suggested that C. papaya fruit extract could be a competitive reducing and stabilizing agent during phytofabrication of nanoparticles. UV–Vis and FTIR spectroscopy showed the formation of SeNPs from sodium selenite, which could be related to the reducing and stabilizing activities of C. papaya fruit extract. The SeNPs were found to be stable with a Zeta potential of −32 mV. The average hydrodynamic size of SeNPs was found as 159 nm by dynamic light scattering. The SeNPs showed a broader XRD pattern with no sharp Bragg's peaks and found to be amorphous. SEM showed that SeNPs were spherical in shape and EDX pattern showed that SeNPs were made up of Se (71.81%), C (11.41%), and O (14.88%). The HR-TEM picture showed that SeNPs were spherical in morphology and have a size range of 101–137 nm. The SeNPs exhibited potent antioxidant activity and their EC50 values (effective concentration required to inhibit 50% of radicals) were 45.65 ± 2.01 and 43.06 ± 3.80 μg/ml in DPPH and ABTS assays, respectively. The antimicrobial action of SeNPs was found as a broad spectrum and suppressed microbial pathogens in ascending order: fungi > Gram-positive bacteria > Gram-negative bacteria. The SeNPs have been demonstrated to reduce the growth and ochratoxin A (OTA) of mycotoxigenic Aspergillus ochraceus and Penicillium verrucosum at 40 μg/ml in broth culture, which is noteworthy. The SeNPs reduced cancer cell proliferation (RAW 264.7, Caco-2, MCF-7, and IMR-32) more preferentially than normal cells (Vero), found to be highly biocompatible. Lower doses of SeNPs (up to 50 μg/ml) were shown to be less toxic and did not cause death in Danio rerio (zebrafish) embryos, implying that lower doses of SeNPs could be beneficial for biological purposes. The present study concluded that phytofabricated SeNPs have multiple biofunctional properties, including antioxidant, antimicrobial, antimycotoxin, and anticancer activities, as well as high biocompatibility. [ABSTRACT FROM AUTHOR]

The study is directed to establish the minimizing effects of Syzygium aromaticum, Ocimum sanctum, and Cananga odorata essential oils on the growth and ochratoxin A (OTA) level of Aspergillus ochraceus and Penicillium verrucosum in maize grains. S. aromaticum essential oil (SAEO), O. sanctum essential oil (OSEO), and C. odorata essential oil (COEO) were extracted by hydro-distillation technique, and a total of 50, 44, and 48 chemical constituents were identified by gas chromatography-mass spectrometry (GC-MS), respectively. The SAEO and OSEO belong to the chemotype of eugenol, whereas, COEO was found to be the chemotype of thymol, limonene, and a-ylangene. The antifungal activity of essential oils (EOs) was determined by the micro-well dilution technique. The SAEO showed superior antifungal activity compared to OSEO, COEO, and synthetic antifungal agent nystatin, and its minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values against A. ochraceous and P. verrucosum were noticed as 1251 ± 42.32 and 1878 ± 28.47 µg/mL, and 0815 ± 22.69 and 1146 ± 51.19 µg/mL, respectively. The antifungal mechanism of EOs was unveiled by assessing the intracellular reactive oxygen species (ROS), ergosterol content, and membrane integrity. The antifungal investigations found that EOs caused fungal mortality by increasing the intracellular ROS, depleting ergosterol synthesis, and distracting membrane integrity. Finally, antifungal and antimycotoxin activity of EOs was demonstrated in maize grains. The SAEO, OSEO, and COEO have reduced the complete fungal growth and OTA level of A. ochraceous and P. verrucosum correspondingly at 2500 and 2500, 3500 and 2500, and 3500 and 3500 µg/g in maize. The EOs could act as natural antifungal agents; protect foodstuffs from fungal infection and mycotoxins during storage. [ABSTRACT FROM AUTHOR]

Aflatoxins (AF) and ochratoxin A (OTA) are fungal metabolites that have carcinogenic, teratogenic, embryotoxic, genotoxic, neurotoxic, and immunosuppressive effects in humans and animals. The increased consumption of plant-based foods and environmental conditions associated with climate change have intensified the risk of mycotoxin intoxication. This study aimed to investigate the abilities of eleven selected LAB strains to reduce/inhibit the growth of Aspergillus flavus, Aspergillus parasiticus, Aspergillus carbonarius, Aspergillus niger, Aspergillus welwitschiae, Aspergillus steynii, Aspergillus westerdijkiae, and Penicillium verrucosum and AF and OTA production under different temperature regiments. Data were treated by ANOVA, and machine learning (ML) models able to predict the growth inhibition percentage were built, and their performance was compared. All factors LAB strain, fungal species, and temperature significantly affected fungal growth and mycotoxin production. The fungal growth inhibition range was 0–100%. Overall, the most sensitive fungi to LAB treatments were P. verrucosum and A. steynii, while the least sensitive were A. niger and A. welwitschiae. The LAB strains with the highest antifungal activity were Pediococcus pentosaceus (strains S11sMM and M9MM5b). The reduction range for AF was 19.0% (aflatoxin B1)-60.8% (aflatoxin B2) and for OTA, 7.3–100%, depending on the bacterial and fungal strains and temperatures. The LAB strains with the highest anti-AF activity were the three strains of P. pentosaceus and Leuconostoc mesenteroides ssp. dextranicum (T2MM3), and those with the highest anti-OTA activity were Leuconostoc paracasei ssp. paracasei (3T3R1) and L. mesenteroides ssp. dextranicum (T2MM3). The best ML methods in predicting fungal growth inhibition were multilayer perceptron neural networks, followed by random forest. Due to anti-fungal and anti-mycotoxin capacity, the LABs strains used in this study could be good candidates as biocontrol agents against aflatoxigenic and ochratoxigenic fungi and AFL and OTA accumulation.

Ochratoxin A (OTA) is one of the most abundant food-contaminating mycotoxins. Besides their high toxicity, mycotoxins are highly stable to physical, chemical or biological detoxification. Therefore, the treatment with cold atmospheric plasma could be one approach to reduce the amount of mycotoxins in different products. The aim of this study was to determine the influence of cold atmospheric plasma on the inactivation of Aspergillus niger and Penicillium verrucosum inoculated on barley and their production of OTA. Inoculated barley was treated with plasma generated by dry air, CO2 or CO2 + O2 for 1 or 3 min and stored for up to two weeks at 9, 25, or 37°C. Three minutes of air plasma treatment effectively significantly reduced the total mold count of both microorganisms by 2.5–3 log cycles. The production of OTA from A. niger was only low, therefore the treatment effect was indistinguishable. The treatment of P. verrucosum on barley after an incubation of five days using a CO2 + O2 plasma resulted in a reduction of the OTA content from 49.0 (untreated) to 27.5 (1 min) and 23.8 ng/g (3 min), respectively. In contrast, CO2 plasma caused an increase of the OTA amount from 49.0 (untreated) to 55.8 (1 min) and 72.9 ng/g (3 min). Finally, the use of air plasma resulted likewise in a decrease of the OTA concentration from 56.9 (untreated) to 25.7 (1 min) and 20.2 ng/g (3 min), respectively. Reducing the incubation time before the treatment to 24 h caused in contrast an increase of the OTA content from 3.1 (untreated) to 29.1 (1 min) and 20.7 ng/g (3 min). Due to the high standard deviation, these changes were not significant, but the tendencies were clearly visible, showing the strong impact of the plasma gas on the OTA production. The results show, that even if the total mold count was reduced, under certain conditions the OTA amount was yet enhanced, probably due to a stress reaction of the mold. Concluding, the plasma gas and incubation conditions have to be considered to allow a successful inactivation of molds and in particular their toxic metabolites.

The present study examined the influence of primary food components on the antifungal activity of the essential oil of Origanum vulgare, carvacrol, thymol, eugenol, and trans-cinnamaldehyde against Penicillium verrucosum and Aspergillus westerdijkiae. The MIC was determined in food model media enriched with proteins (1, 5, or 10%), carbohydrates (1, 4, or 6%), or oil (1, 5, or 10%). Proteins increased the antifungal activity of O. vulgare essential oil, carvacrol, thymol, and eugenol, whereas the effect of trans-cinnamaldehyde decreased with increasing protein content. The presence of carbohydrates diminished the inhibitory effect of the natural preservatives on A. westerdijkiae; for P. verrucosum, their inhibitory effect increased with carbohydrates. Only the antifungal activity of trans-cinnamaldehyde did not depend on the carbohydrate content. The presence of oil had the strongest influence. At a concentration of 1% oil, the antifungal activity decreased significantly, and at 10% oil, almost no inhibition was observed. To investigate the effect of the antifungal agents on the morphology of the target molds, they were grown on malt extract agar containing carvacrol and trans-cinnamaldehyde and were examined by scanning electron microscopy. The hyphae, conidiophores, vesicles, and phialides were severely altered and deformed, and spore formation was clearly suppressed. The antifungal activity of natural preservatives is influenced by the food matrix. Proteins increase the impact of O. vulgare EO, carvacrol, thymol, and eugenol. Carbohydrates diminish the inhibition of natural preservatives on A. westerdijkiae. In the presence of oil, natural inhibitors lose their antifungal effect. Carvacrol and trans-cinnamaldehyde lead to distorted hyphae and loss of sporulation. [ABSTRACT FROM AUTHOR]

The present study examines the influence of the natural preservatives carvacrol, eugenol, trans-cinnamaldehyde and the essential oil (EO) Origanum vulgare on ochratoxin A (OTA) production and the mycelial growth of two food-related moulds, Penicillium verrucosum and Aspergillus westerdijkiae, by broth macro-dilution assay for 21 days. With the addition of ½ minimum inhibitory concentration (MIC) carvacrol, eugenol and O. vulgare EO, the mycelial dry weight of both moulds decreased significantly over the whole incubation period of 7, 14 and 21 days. Trans-cinnamaldehyde slightly stimulated the growth of A. westerdijkiae and P. verrucosum at 14 and 21 days of incubation. Growth inhibition did not accompany inhibition of OTA production. Although the growth of both moulds was inhibited after the addition of ½ MIC carvacrol, eugenol and O. vulgare EO, the OTA production of the strong mycotoxin producer A. westerdijkiae was stimulated. Only trans-cinnamaldehyde inhibited the production of OTA with the addition of ½ MIC. P. verrucosum produced significantly less OTA than A. westerdijkiae, and its mycotoxin production was almost completely inhibited by the addition of ½ MIC of the natural preservatives. [ABSTRACT FROM AUTHOR]

Background and objectives: Ochratoxin A (OTA) is one of the most prevalent mycotoxins in cereal grains. In temperate climates, Penicillium verrucosum is the primary OTA producer. Our objectives were (a) to determine the effects of various factors on OTA accumulation in oat grain inoculated with P. verrucosum and (b) to develop a method to screen oat cultivars for OTA accumulation. Findings: Grain samples from three oat cultivars were inoculated with three different isolates of P. verrucosum, incubated at two water activity levels (0.85 and 0.90 aw) and two temperatures (22.5 and 27.5°C) for 4, 7, and 10 weeks. A water activity of 0.90 and a temperature of 22.5°C provided the highest level of OTA accumulation. Isolate C1136‐1 was the highest OTA producer among the isolates of P. verrucosum studied. The longer the incubation period, the higher the OTA accumulation in oat grain samples was. Conclusions: The P. verrucosum isolate, the water activity and temperature during incubation, and the incubation period, all had an effect on the level of OTA accumulation. Inoculating grains with isolate C84‐2 and incubating at 22.5°C and 0.90 aw for 7 weeks provided adequate conditions to screen oat cultivars for OTA accumulation. Significance and novelty: A new protocol was developed that can be used for discriminating among oat cultivars for level of OTA accumulation. It would ultimately help breeders and pathologists in selecting genotypes with reduced OTA accumulation. [ABSTRACT FROM AUTHOR]

Ecological conditions during ripening of dry-cured meat products favour the growth of an uncontrolled mould population that could suppose a risk of ochratoxin A (OTA) production. In this work the influence of water activity (a w), temperature and substrate composition on fungal growth and OTA production by Penicillium nordicum and Penicillium verrucosum isolated from dry-cured meat products have been studied. In addition, the relative risk of OTA presence on dry-cured meat products has been evaluated using the Risk Ranger software. Fungal growth was observed in the range of 0.99–0.90 a w and 15–25 °C being mainly temperature-dependent. P. nordicum and P. verrucosum were able to produce OTA in every substrate in these ranges of a w and temperature. The production of OTA by P. verrucosum was mainly influenced by temperature and media composition. However, P. nordicum it is affected mainly by substrate or temperature depending on the strain studied. Both species produce a large amount of OTA on dry-cured ham and on dry-cured fermented sausage "salchichón" in environmental conditions usually found throughout the ripening of these products. The Risk Ranger software reveals that the relative risk of OTA on dry-cured meat products is 75%. Thus, control measures during dry-cured meat products processing to prevent OTA risk should be established. Unlabelled Image [ABSTRACT FROM AUTHOR]

The aims of this study were to evaluate the antifungal activity of the bioactive compound allyl isothiocyanate (AITC) against Aspergillus flavus (8111 ISPA) aflatoxins (AFs) producer and Penicillium verrucosum (D-01847 VTT) ochratoxin A (OTA) producer on corn, barley, and wheat. The experiments were carried out initially in a simulated silo system for laboratory scale composed of glass jars (1 L). Barley and wheat were contaminated with P. verrucosum and corn with A. flavus. The cereals were treated with a hydroxyethylcellulose gel disk to which 500 µL/L of AITC were added; the silo system was closed and incubated for 30 days at 21 °C. After that, simulated silos of 100 L capacity were used. Barley, wheat, and corn were contaminated under the same conditions as the previous trial and treated with disks with 5 mL of AITC, closed and incubated for 90 days at 21 °C. In both cases, the control test did not receive any antifungal treatment. The growth of the inoculated fungi and the reduction in the formation of AFs and OTA were determined. In the lab scale silo system, complete inhibition of fungal growth at 30 days has been observed. In corn, the reduction of aflatoxin B1 (AFB1) was 98.5%. In the 100 L plastic drums, a significant reduction in the growth of A. flavus was observed, as well as the OTA formation in wheat (99.5%) and barley (92.0%). [ABSTRACT FROM AUTHOR]

Background and objectives: Ochratoxin A (OTA) is a carcinogenic and nephrotoxic mycotoxin produced by various Penicillium and Aspergillus species. Cereal grains, including oats, are among the food sources that can be contaminated with OTA. Grains of twelve oat cultivars grown in seven environments were inoculated with P. verrucosum and incubated at 0.90 aw and 22.5°C for 7 weeks. Our objectives were to (a) determine the effect of cultivar and growing environment on OTA contamination in oat grain and (b) identify oat grain characteristics associated with OTA accumulation. Findings: The effect of the cultivar was significant in two of the environments considered. When data from all seven environments were combined for analysis, significant differences among oat cultivars were also observed. Grain composition and grain physical characteristics were determined to identify characteristics associated with OTA accumulation. We found that test weight, groat percent, the proportion of plump groats, and groat oil content were significantly correlated with the level of OTA accumulation in the grains. Conclusions: Significant differences among genotypes for level of OTA accumulation suggest that cultivars with lower levels of OTA accumulation can be used to help mitigate risks of OTA contamination in oat grain. Significance and novelty: This study is the first to report differences among oat cultivars for OTA accumulation. [ABSTRACT FROM AUTHOR]

Abstract Ochratoxigenic penicillia can grow on the cured meat products surface during their ripening. Ingredients added to cured meat products throughout their processing, and specifically of dry-fermented sausages, consist of a potential strategy to prevent or minimise the hazard associated with ochratoxin A (OTA) in these products. The aim of this work was to evaluate the effect of different concentrations of NaCl, KCl and sucrose in a conducive medium on the growth and OTA production by two Penicillium verrucosum strains. In general, there were no statistical differences between their growth rates. Although OTA production was not completely avoided in the presence of KCl and sucrose, it was reduced by comparison with NaCl. The concentration of the ingredients also affected the OTA production by P. verrucosum. These results would mean that the replacement of NaCl with KCl would imply the production of potentially safer cured meat products in terms of the OTA presence, which is of great importance for the current trends to reduce the NaCl levels in this kind of products. Furthermore, the addition of sucrose should be considered one of the approaches of the hurdle technology for minimising OTA accumulation in cured meat products. Highlights • The role of cured meat ingredients on mould growth and OTA production was evaluated. • Two ochratoxigenic Penicillium verrucosum strains were investigated. • Type and amount of cured meat ingredients influenced growth and OTA accumulation. • Large differences between the tested two P. verrucosum strains were not found. • Formulation changes might provide safer cured meats in terms of the OTA presence. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is one of the worldwide most important mycotoxins in terms of health and agroeconomic consequences. With the aim to promote the use of phytochemicals as alternatives to synthetic fungicides, the effect of hydroxycinnamic acids on the fungal growth and OTA yield by two major OTA-producing species was investigated. After a first step dedicated to the definition of most suitable culture conditions, the impact of 0.5 mM ferulic (FER), p-coumaric (COUM), caffeic and chlorogenic acids was evaluated on Aspergillus westerdijkiae and Penicillium verrucosum. Whereas no fungal growth reduction was observed regardless of the phenolic acid and fungal isolate, our results demonstrated the capacity of FER and COUM to inhibit OTA production. The most efficient compound was FER that led to a 70% reduction of OTA yielded by P. verrucosum and, although not statistically significant, a 35% inhibition of OTA produced by A. westerdijkiae. To further investigate the bioactivity of FER and COUM, their metabolic fate was characterized in fungal broths. The capacity of P. verrucosum to metabolize FER and COUM through a C2-clivage type degradation was demonstrated. Overall, our data support the potential use of FER to prevent OTA contamination and reduce the use of synthetic pesticides.

During the last decades, essential oils (EOs) have been proven to be a natural alternative to additives or pasteurization for the prevention of microbial spoilage in several food matrices. In this work, we tested the antimicrobial activity of EOs from Melissa officinalis, Ocimum basilicum, and Thymus vulgaris against three different microorganisms: Escherichia coli, Clostridium tyrobutyricum, and Penicillium verrucosum. Pressed ewes’ cheese made from milk fortified with EOs (250 mg/kg) was used as a model. The carryover effect of each oil was studied by analyzing the volatile fraction of dairy samples along the cheese-making process using headspace stir bar sorptive extraction coupled to gas chromatography/mass spectrometry. Results showed that the EOs contained in T. vulgaris effectively reduced the counts of C. tyrobutyricum and inhibited completely the growth of P. verrucosum without affecting the natural flora present in the cheese. By contrast, the inhibitory effect of M. officinalis against lactic acid bacteria starter cultures rendered this oil unsuitable for this matrix.

Ochratoxin A (OTA) can cause toxicogenic effects in humans and animals when contaminated food products are consumed. Oat (Avena sativa), like any other cereal grain, can be contaminated with OTA when storage conditions are favorable for fungal growth and toxin production. South Dakota is among the leading oat-producing states in the United States. It is therefore important to determine the frequency of occurrence of the primary OTA-producing fungal species on oat grains produced in the state. In this study, we evaluated oat grain samples from South Dakota for the incidence of Penicillium verrucosum, the major ochratoxigenic fungus in temperate regions. Kernels from 12 oat cultivars grown at multiple locations in South Dakota from 2014 to 2016 (15 location-year combinations) were plated on dichloran yeast extract sucrose glycerol agar medium. P. verrucosum was detected on 0.30, 0.19, and 0.05% of the kernels tested in 2014, 2015, and 2016, respectively. Overall, 22 of the 360 evaluated samples had kernels contaminated with P. verrucosum. The fungal incidence of the contaminated samples ranged from 1 to 16%, and the majority of those samples originated from one location. All samples from 2014 and 2015 (n=240), except two, had no detectable levels of OTA. The concentration of OTA was well under the maximum limit recommended by the European Union for the two samples with detectable levels of OTA. [ABSTRACT FROM AUTHOR]

This study deals with the efficacy of Origanum onites, Salvia officinalis, and Mentha piperita essential oils (EOs) and their mixture (1:1:1, v/v/v) against ochratoxigenic P. verrucosum D-99756 and modelling the effects of EOs on fungal growth using modified Gomperz and logistic models. O. onites, S. officinalis, and M. piperita EOs were used at their minimal lethal concentration levels at 65, 125, and 250 μL/mL, respectively, which were determined in our preliminary studies. Inactivation of fungi sporulation by EO treatments was higher than 50-60% within 48 h, while 78.50-81.41% lethality was observed at the end of the 168 h for all of the tested EOs. The most effectiveness time for lethal effect was found at 84 h for O. onites and S. officinalis, while it was 168 h for mix EOs (p < 0.05). The modified Gompertz model successfully produced better fits than the logistic model as evidenced by the root mean square error, the modelling efficiency, and accuracy efficiency. In conclusion, data confirmed the potential use of plant and spice EOs for their fungistatic and fungicidal effects in food systems. [ABSTRACT FROM AUTHOR]

The aims of this study were to evaluate the antifungal activity of the bioactive compound allyl isothiocyanate (AITC) against Aspergillus flavus (8111 ISPA) aflatoxins (AFs) producer and Penicillium verrucosum (D-01847 VTT) ochratoxin A (OTA) producer on corn, barley, and wheat. The experiments were carried out initially in a simulated silo system for laboratory scale composed of glass jars (1 L). Barley and wheat were contaminated with P. verrucosum and corn with A. flavus. The cereals were treated with a hydroxyethylcellulose gel disk to which 500 µL/L of AITC were added; the silo system was closed and incubated for 30 days at 21 °C. After that, simulated silos of 100 L capacity were used. Barley, wheat, and corn were contaminated under the same conditions as the previous trial and treated with disks with 5 mL of AITC, closed and incubated for 90 days at 21 °C. In both cases, the control test did not receive any antifungal treatment. The growth of the inoculated fungi and the reduction in the formation of AFs and OTA were determined. In the lab scale silo system, complete inhibition of fungal growth at 30 days has been observed. In corn, the reduction of aflatoxin B1 (AFB1) was 98.5%. In the 100 L plastic drums, a significant reduction in the growth of A. flavus was observed, as well as the OTA formation in wheat (99.5%) and barley (92.0%).

The aim of this study was to investigate the effect of synthetic detergent Merix (Henkel, Kruševac, Serbia), and its particular components-ethoxylated oleyl-cetyl alcohol and sodium tripolyphosphate on the growth and metabolic activity of Penicillium verrucosum. During 19 days of fungal cultivation in Czapek-Dox liquid medium supplemented with or without 0.5% pollutants, the following parameters were observed: pH, the total biomass dry weight, the quantity of free and total organic acids and proteolytic activity. The detergent caused a slight stimulatory (2.41%) effect whereas sodium tripolyphosphate and ethoxylated oleyl-cetyl alcohol provided a slight inhibitory action (0.59 and 2.75%, respectively) on the fungal biomass. The pollutants decreased pH values of the media and the quantity of free organic acids. In contrast, they enhanced the quantity of total organic acids. Proteolytic activity remained nearly unchanged (95.8%) in the presence of detergent and reduced to 80.1% in sodium tripolyphosphate-supplemented medium. In contrast, the enzyme activity sharply increased (260.8%) with ethoxylated oleyl-cetyl alcohol. The obtained results indicate the potential of P. verrucosum in bioremediation of environment contaminated with synthetic detergent taken in high concentration. [ABSTRACT FROM AUTHOR]

Penicillium verrucosum, P. nordicum and Aspergillus carbonarius are three important ochratoxin A producing species. P. verrucosum is in addition able to produce citrinin. It has been shown earlier that P. nordicum is adapted to NaCl rich environments like salt rich dry cured foods or even salines. In this organism, the biosynthesis of ochratoxin A plays an adaptive role in this habitat. P. verrucosum generally can be found on cereals, but occasionally also on salt rich dry cured foods. In contrast A. carbonarius usually cannot be found in NaCl rich environments, but it occurs in another environment with high concentration of solutes, e.g., in sugar rich substrates like grapes and grape juices. Usually osmotic challenging conditions activate the HOG MAP kinase signal cascade, which in turn activates various osmo-regulated genes. In the current analysis, it could be demonstrated that in case of P. nordicum and P. verrucosum the NaCl induced production of ochratoxin A is correlated to the phosphorylation status of the HOG MAP kinase. Just the opposite was true for A. carbonarius. In this case, also higher amounts of NaCl in the medium lead to an increased phosphorylation status of HOG, but no increase in ochratoxin biosynthesis was observed. In contrast to the Penicillia, higher NaCl concentrations lead to a rapid cessation of growth by A. carbonarius. High glucose concentrations have much less impact on growth and the phosphorylation of HOG.