The characteristics, occurrence, and toxicological effects of alternariol: a mycotoxin.

Alternaria species are mycotoxin-producing fungi known to infect fresh produce and to cause their spoilage. Humans get exposed to fungal secondary metabolites known as mycotoxin via the ingestion of contaminated food. Alternariol (AOH) (C14H10O5) is an isocoumarins produced by different species of Alternaria including Alternaria alternata. AOH is often found in grain, fruits and fruits-based food products with high levels in legumes, nuts, and tomatoes. AOH was first discovered in 1953, and it is nowadays linked to esophagus cancer and endocrine disruption due to its similarity to estrogen. Although considered as an emerging mycotoxin with no regulated levels in food, AOH occurs in highly consumed dietary products and has been detected in various masked forms, which adds to its occurrence. Therefore, this comprehensive review was developed to give an overview on recent literature in the field of AOH. The current study summarizes published data on occurrence levels of AOH in different food products in the last ten years and evaluates those levels in comparison to recommended levels by the regulating entities. Such surveillance facilitates the work of health risk assessors and highlights commodities that are most in need of AOH levels regulation. In addition, the effects of AOH on cells and animal models were summarized in two tables; data include the last two-year literature studies. The review addresses also the main characteristics of AOH and the possible human exposure routes, the populations at risk, and the effect of anthropogenic activities on the widespread of the mycotoxin. The commonly used detection and control methods described in the latest literature are also discussed to guide future researchers to focus on mitigating mycotoxins contamination in the food industry. This review aims mainly to serve as a guideline on AOH for mycotoxin regulation developers and health risk assessors.

Occurrence of Mycotoxins and Toxigenic Fungi in Cereals and Application of Yeast Volatiles for Their Biological Control.

Fungal infections in cereals lead to huge economic losses in the food and agriculture industries. This study was designed to investigate the occurrence of toxigenic fungi and their mycotoxins in marketed cereals and explore the effect of the antagonistic yeast Cyberlindnera jadinii volatiles against key toxigenic fungal strains. Aspergillus spp. were the most frequent contaminating fungi in the cereals, with an isolation frequency (Fr) of 100% in maize, followed by wheat (88.23%), rice (78.57%) and oats (14.28%). Morphological and molecular identification confirmed the presence of key toxigenic fungal strains in cereal samples, including A. carbonarius, A. flavus, A. niger, A. ochraceus and A. parasiticus. Aflatoxins (AFs) were detected in all types of tested cereal samples, with a significantly higher level in maize compared to wheat, rice, oats and breakfast cereals. Ochratoxin A (OTA) was only detected in wheat, rice and maize samples. Levels of mycotoxins in cereals were within EU permissible limits. The volatiles of Cyberlindnera jadinii significantly inhibited the growth of A. parasiticus, A. niger and P. verrucosum. The findings of this study confirm the presence of toxigenic fungi and mycotoxins in cereals within the EU permissible limits and the significant biocontrol ability of Cyberlindnera jadinii against these toxigenic fungi.

Detection of multimycotoxins in camel feed and milk samples and their comparison with the levels in cow milk.

Camel milk has been considered as an important source of nutrients and is commercialized in many countries of the world including the Middle East. This study aimed to investigate the presence of mycotoxins in camel feed and milk samples in comparison with the cow milk. Fumonisins (FUM), ochratoxin A (OTA), and zearalenone (ZEN) were detected in 14%, 39%, and 39% of the tested camel feed samples, respectively. Among the tested camel feed samples, 8.3% and 5.6% were co-contaminated with OTA+FUM and FUM+ZEN, respectively. In the case of milk samples, 46.15% of camel and 63.63% of cow were found contaminated with aflatoxin M1 (AFM1). In total, 16.2% and 8.1% of the milk samples were simultaneously contaminated with two and three mycotoxins, respectively. Although the levels of individual mycotoxins in the camel feed and milk samples were within the European Union (EU) permissible limits, their co-occurrence may pose severe risk to human and animal health due to possible additive and/or synergistic toxicities.

Biocontrol Activity of Bacillus megaterium BM344-1 against Toxigenic Fungi.

Mycotoxins are secondary metabolites of some fungal species and represent important contaminants of food and feed. This study aimed to explore the biological control activity of Bacillus megaterium BM344-1 volatile organic compounds (VOCs) on the growth and mycotoxin production of single representatives of the toxigenic species Aspergillus flavus, Aspergillus carbonarius, Penicillium verrucosum, and Fusarium verticillioides. In vitro co-incubation experiments indicated the P. verrucosum isolate as the most sensitive one, with a growth inhibition ratio of 66.7%, followed by A. flavus (29.4%) and F. verticillioides (18.2%). Exposure of A. flavus, P. verrucosum, and F. verticillioides to BM344-1 VOCs resulted in complete inhibition of aflatoxins (AFB1, AFG1, and AFG2), ochratoxin A, and fumonisin B1 (FB1) synthesis on artificial media, respectively. In vivo experiments on maize kernels showed 51% inhibition of fungal growth on ears simultaneously infected with A. flavus spores and exposed to BM344-1 volatiles. Likewise, AF synthesis by A. flavus was significantly (p < 0.05) inhibited (25.34 ± 6.72 µg/kg) by bacterial volatiles as compared to that in control maize ears (91.81 ± 29.10 µg/kg). Gas chromatography-tandem mass spectrometry-based analysis of headspace volatiles revealed hexadecanoic acid methyl ester (palmitic acid) and tetracosane as bioactive compounds in the BM344-1 volatilome. Bacterial volatiles have promising potential to control the growth and mycotoxin synthesis of toxigenic fungi and may present valuable aid in the efforts to warrant food and feed safety.

Isolation of a Novel Kluyveromyces marxianus Strain QKM-4 and Evidence of Its Volatilome Production and Binding Potentialities in the Biocontrol of Toxigenic Fungi and Their Mycotoxins.

To overcome the economic losses associated with fungi and their toxic metabolites, environmentally safe and efficient approaches are needed. To this end, biological control using yeasts and safe bacterial strains and their products are being explored to replace synthetic fungicides. In the present study, the biocontrol effect of a yeast strain of Kluyveromyces marxianus, QKM-4, against the growth and mycotoxin synthesis potential of key toxigenic fungi was evaluated. In vitro assays were performed to find the application of yeast volatile organic compounds (VOCs) against fungal contamination on important agricultural commodities. The removal of ochratoxin A (OTA) and deoxynivalenol (DON) by living and heat-inactivated yeast cells was also explored. VOCs produced by strain QKM-4 were able to significantly limit the fungal growth of 17 fungal species belonging to genera Aspergillus, Penicillium, and Fusarium. Yeast VOCs were able to reduce OTA biosynthesis potential of Penicillium verrucosum and Aspergillus carbonarius by 99.6 and 98.7%, respectively. In vivo application of QKM-4 VOCs against Fusarium oxysporum and A. carbonarius infection on tomatoes and grapes, respectively, determined a complete inhibition of fungal spore germination. GC/MS-based analysis of yeast VOCs identified long-chain alkanes, including nonadecane, eicosane, docosane, heptacosane, hexatriacontane, and tetracosane. In vitro testing of the mycotoxin-binding potential of the living and heat-inactivated QKM-4 cells showed a reduction of OTA and DON up to 58 and 49%, respectively, from artificially contaminated buffers. Our findings clearly demonstrate the strong antifungal potential of K. marxianus QKM-4 and propose this strain as a strong candidate for application in agriculture to safeguard food and feed products.

In-vitro Application of a Qatari Burkholderia cepacia strain (QBC03) in the Biocontrol of Mycotoxigenic Fungi and in the Reduction of Ochratoxin A biosynthesis by Aspergillus carbonarius.

Mycotoxins are secondary metabolites produced by certain filamentous fungi, causing human and animal health issues upon the ingestion of contaminated food and feed. Among the safest approaches to the control of mycotoxigenic fungi and mycotoxin detoxification is the application of microbial biocontrol agents. Burkholderiacepacia is known for producing metabolites active against a broad number of pathogenic fungi. In this study, the antifungal potential of a Qatari strain of Burkholderia cepacia (QBC03) was explored. QBC03 exhibited antifungal activity against a wide range of mycotoxigenic, as well as phytopathogenic, fungal genera and species. The QBC03 culture supernatant significantly inhibited the growth of Aspergillus carbonarius, Fusarium culmorum and Penicillium verrucosum in PDA medium, as well as A. carbonarius and P. verrucosum biomass in PDB medium. The QBC03 culture supernatant was found to dramatically reduce the synthesis of ochratoxin A (OTA) by A. carbonarius, in addition to inducing mycelia malformation. The antifungal activity of QBC03's culture extract was retained following thermal treatment at 100 °C for 30 min. The findings of the present study advocate that QBC03 is a suitable biocontrol agent against toxigenic fungi, due to the inhibitory activity of its thermostable metabolites.

Application of Low-Fermenting Yeast Lachancea thermotolerans for the Control of Toxigenic Fungi Aspergillus parasiticus, Penicillium verrucosum and Fusarium graminearum and Their Mycotoxins.

Mycotoxins are important contaminants of food and feed. In this study, low fermenting yeast (Lachancea thermotolerans) and its derivatives were applied against toxigenic fungi and their mycotoxins. A. parasiticus, P. verrucosum and F. graminearum and their mycotoxins were exposed to yeast volatile organic compounds (VOCs) and cells, respectively. VOCs reduced significantly the fungal growth (up to 48%) and the sporulation and mycotoxin synthesis (up to 96%). Very interestingly, it was shown that even 7 yeast colonies reduced Fusarium’s growth and the synthesis of its mycotoxin, deoxynivalenol (DON). Moreover, decreasing yeast nutrient concentrations did not affect the inhibition of fungal growth, but reduced DON synthesis. In addition, inactivated yeast cells were able to remove up to 82% of the ochratoxin A (OTA). As an application of these findings, the potentialities of the VOCs to protect tomatoes inoculated with F. oxysporum was explored and showed that while in the presence of VOCs, no growth was observed of F. oxysporum on the inoculated surface areas of tomatoes, in the absence of VOCs, F. oxysporum infection reached up to 76% of the tomatoes’ surface areas. These results demonstrate that the application of yeasts and their derivatives in the agriculture and food industry might be considered as a very promising and safe biocontrol approach for food contamination.