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Aflatoxins (AFs) are hazardous carcinogens and mutagens produced by some molds, particularly Aspergillus spp. Therefore, the purpose of this study was to isolate and identify endophytic bacteria, extract and characterize their bioactive metabolites, and evaluate their antifungal, antiaflatoxigenic, and cytotoxic efficacy against brine shrimp (Artemia salina) and hepatocellular carcinoma (HepG2). Among the 36 bacterial strains isolated, ten bacterial isolates showed high antifungal activity, and thus were identified using biochemical parameters and MALDI-TOF MS. Bioactive metabolites were extracted from two bacterial isolates, and studied for their antifungal activity. The bioactive metabolites (No. 4, and 5) extracted from Bacillus cereus DSM 31T DSM, exhibited strong antifungal capabilities, and generated volatile organic compounds (VOCs) and polyphenols. The major VOCs were butanoic acid, 2-methyl, and 9,12-Octadecadienoic acid (Z,Z) in extracts No. 4, and 5 respectively. Cinnamic acid and 3,4-dihydroxybenzoic acid were the most abundant phenolic acids in extracts No. 4, and 5 respectively. These bioactive metabolites had antifungal efficiency against A. flavus and caused morphological alterations in fungal conidiophores and conidiospores. Data also indicated that both extracts No. 4, and 5 reduced AFB1 production by 99.98%. On assessing the toxicity of bioactive metabolites on A. salina the IC50 recorded 275 and 300 µg/mL, for extracts No. 4, and 5 respectively. Meanwhile, the effect of these extracts on HepG2 revealed that the IC50 of extract No. 5 recorded 79.4 µg/mL, whereas No. 4 showed no cytotoxic activity. It could be concluded that bioactive metabolites derived from Bacillus species showed antifungal and anti-aflatoxigenic activities, indicating their potential use in food safety.

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Aflatoxins are an unavoidable contaminant of foods. The current work aimed to study the ameliorating effect of Lawsonia inermis L. extract and its nano-formulation versus aflatoxin ingestion in ulcerative rats. Lawsonia inermis L. bioactivity was evaluated by both antioxidant & antimicrobial assays. The nanoparticles characterization measurements were evaluated. Different parameters in the fortified milk beverage were assessed. Seventy two Sprague-Dawley male rats were randomized into 12 groups (6 rats/group) where peptic ulcer was induced with a single aspirin dose (500 mg/kg BW) orally. The nutritional and biochemical parameters were evaluated. The results showed that antioxidant activity and total phenolic content increased with increasing nano-formulation ratio. A remarkable improvements in all the treated groups, either for ulcer alone or for aflatoxin exposed ulcerative groups in normal and nano-formulation. Conclusively, Lawsonia inermis L. & its nano-formulation could act as dual therapy for ulcer treatment and the hazardous effects of aflatoxin exposure.

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Aflatoxins are toxic carcinogens and mutagens formed by some moulds, specifically Aspergillus spp. Therefore, this study aimed to extract and identify bioactive secondary metabolites from Lactobacillus species, to evaluate their efficacy in reducing fungal growth and aflatoxin production and to investigate their toxicity. The bioactive secondary metabolites of Lactobacillus species showed variable degrees of antifungal activity, whereas L. rhamnosus ethyl acetate extract No. 5 exhibited the highest antifungal activity and, thus, was selected for further identification studies. Data revealed that L. rhamnosus ethyl acetate extract No. 5 produced various organic acids, volatile organic compounds and polyphenols, displayed antifungal activity against A. flavus, and triggered morphological changes in fungal conidiophores and conidiospores. L. rhamnosus ethyl acetate extract No. 5 at a 9 mg/mL concentration reduced AFB1 production by 99.98%. When the effect of L. rhamnosus ethyl acetate extract No. 5 on brine shrimp mortality was studied, the extract attained a 100% mortality at a concentration of 400 µg/mL, with an IC50 of 230 µg/mL. Meanwhile, a mouse bioassay was performed to assess the toxicity of L. rhamnosus ethyl acetate extract No. 5, whereas there were no harmful effects or symptoms in mice injected with L. rhamnosus ethyl acetate extract at concentrations of 1, 3, 5, 7, and 9 mg/kg body weight.

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Chlorpyrifos is an organophosphate insecticide linked to neurological dysfunctions, endocrine disturbance, cardiovascular illness, genotoxicity, histopathological abnormalities, immunotoxicity, and oxidative stress. Therefore, the aim of this study was to prepare activated carbon from agricultural waste to adsorb and remove chlorpyrifos from aqueous solutions, as well as to study the physicochemical characteristics of the prepared activated carbon.Activated carbon was prepared from agricultural waste (banana peels, orange peels, pomegranate peels and date stones). The activated carbon prepared showed an exterior surface that was irregular and full of cavities with Brunauer-Emmett-Teller(BET) surface areas of 94.26, 111.75, 183.89, and 289.86 m2/g for activated carbon prepared from orange peels, date stone, pomegranate peels, and banana peels respectively. The Scanning Electron Microscope (SEM) image revealed that the activated carbon's exterior surface was irregular and full of various shapes and sizes of cavities.The Energy Dispersive X-Ray (EDX) indicated the existence of carbon, oxygen, silicon and potassium in banana peels-derived activated carbon, whereas carbon, oxygen, silicon and potassium, in addition to aluminium, were detected in the pomegranate peels-derived activated carbon. The Fourier-Transform Infrared Spectroscopy (FTIR) analysis of prepared activated carbon revealed several functional groups, including carboxylic acid, carbon dioxide, and aromatic compounds. Results also showed that the activated carbon significantly removed chlorpyrifos from water, recording 97.6%, 90.6%, 71.48%, and 52.00 % for activated carbon prepared from pomegranate peels, banana peels, date stones and orange peels, respectively. The study concluded that agricultural waste-derived activated carbon could be employed as an alternative pesticide adsorbent.

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Organophosphorus pesticides (OPPs) cause great risk to human health as they are used globally. Therefore, the purpose of this research was to determine the total phenolics, flavonoids, and antioxidant activity of agricultural waste, as well as to control the pesticide residues (diazinon, and parathion) at a laboratory scale level using dried-milled fruit wastes. The pesticide residues parathion and diazinon were used at concentrations of 0.094, and 1.90 mg/mL respectively. The fruit wastes used in this study were orange and banana peels, as well as date stones, and they were used in two concentrations (3 and 9 g/30 mL deionized water). The total phenolic and flavonoid contents and the antioxidant activity were measured in fruit wastes. Also, the Fourier transmitted infrared (FTIR) spectra of fruit wastes were established to figure out the nature of the functional groups found before and after pesticide residues removal. The ability of fruit wastes to remove pesticides residues was determined using Gas Chromatography/Mass spectrometry (GC/MS). Data showed that date stones contained a higher amount of total phenolic content than orange and banana peels. However, orange peels contained a higher amount of total flavonoid contents than those of date stones and banana peels. As for antioxidant activity, banana peels recorded the higher antioxidant activity, followed by orange peels and date stones respectively. Results revealed that there was no relation between total phenolic content, total flavonoid content, and antioxidant activity. Results also indicated that date stones at a concentration of 9 g successfully reduced diazinon (81.18%), followed by banana (63.86%) and orange peels (43.42%) respectively, whereas parathion was reduced by banana peels at a concentration of 9 g (50.34%), followed by orange peels (45.28%), and date stones (39.52%) respectively. This study demonstrated that agricultural wastes were effective in the adsorption of diazinon from water, and their use is considered safe for the environment.

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This study aimed to assess the protective effect of encapsulating humic acid-iron complexed nanoparticles (HA-Fe NPs) inside glucanmannan lipid particles (GMLPs) extracted from yeast cell wall against aflatoxin B (AFB1 ) toxicity in vivo. Four groups of male Sprague-Dawley rats were treated orally for 2 weeks included the control group, AFB1 treated group (80 µg/kg b.w); GMLP/HA-Fe NPs treated group (0.5 mg/kg b.w), and the group treated with AFB1 plus GMLP/HA-Fe NPs. GMLPs are empty 3-4 micron permeable microspheres that provide an efficient system for the synthesis and encapsulation of AFB1 -absorbing nanoparticles (NPs). Humic acid nanoparticles (HA-NPs) were incorporated inside the GMLP cavity by complexation with ferric chloride. In vivo study revealed that AFB1 significantly elevated serum alanine aminotransferase, aspartate aminotransferase, creatinine, uric acid, urea, cholesterol, triglycerides, LDL, malondialdehyde, and nitric oxide. It significantly decreased total protein, high-density lipoprotein, hepatic and renal CAT and glutathione peroxidase content and induced histological changes in the liver and kidney (p ≤ 0.05). The coadministration of the synthesized formulation GMLP/HA-Fe NPs with AFB1 has a protective effect against AFB1 -induced hepato-nephrotoxicity, oxidative stress and histological alterations in the liver and kidney.

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Aflatoxins and ochratoxin A occur frequently in grains and are associated with carcinogenic, and nephrotoxic properties. Therefore, the aim of this study was to determine the level of aflatoxins and ochratoxin A contamination in wheat samples obtained from different Egyptian governorates and to assess the effect of gamma irradiation on AFB1 in spiked wheat samples, as well as to evaluate the estimated daily intake and hazard index. Thirty-six wheat grain samples purchased from different sale points were analyzed by High-Performance Liquid Chromatography. Data revealed that 33.33 % of the wheat grain samples were contaminated by aflatoxin B1, whereas only 16.66 % of the wheat samples were above the maximum limits (2 µg/kg) set by the European Commission. Ochratoxin A was only detected in two wheat grain samples, and the results were considered below the maximum limit (5 µg/kg) set by the European Commission. On studying the effect of gamma irradiation on wheat samples spiked by aflatoxin B1 (20 µg/kg), results revealed that aflatoxin B1 was reduced to 1.22 and 0.94 µg/kg for samples gamma-irradiated at a dose of 10 and 20 KGy respectively. Estimated daily intake of ochratoxin A in wheat samples was found to be higher than that of the tolerable daily intake; however, hazard index values were below one, thus demonstrating no threat to human health.

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This study aimed to identify the bioactive compounds of the ethyl acetate extract of Aspergillus niger SH2-EGY using GC-MS and to evaluate their protective role against aflatoxin B1 (AFB1)-induced oxidative stress, genotoxicity and cytotoxicity in rats. Six groups of male Sprague-Dawley rats were treated orally for 4 weeks included the control group, AFB1-treated group (80 µg/kg b.w); fungal extract (FE)-treated groups at low (140) or high dose (280) mg/kg b.w and the groups treated with AFB1 plus FE at the two tested doses. The GC-MS analysis identified 26 compounds. The major compounds found were 1,2,3,4,6-Penta-trimethylsilyl Glucopyranose, Fmoc-L-3-(2-Naphthyl)-alanine, D-(-)-Fructopyranose, pentakis (trimethylsilyl) ether, bis (2-ethylhexyl) phthalate, trimethylsilyl ether-glucitol, and octadecanamide, N-(2- methylpropyl)-N-nitroso. The in vivo results showed that AFB1 significantly increased serum ALT, AST, creatinine, uric acid, urea, cholesterol, triglycerides, LDL, carcinoembryonic antigen, alpha-fetoprotein, interleukin-6, Malondialdehyde, nitric oxide, Bax, caspase-3 and P53 mRNA expression, chromosomal aberrations and DNA fragmentation. It decreased serum TP, albumin, HDL, Bcl-2 mRNA expression, hepatic and renal TAC, SOD and GPx content and induced histological changes in the liver and kidney. FE prevented these disturbances in a dosage-dependent manner. It could be concluded that A. niger SH2-EGY extract is safe a promising agent for pharmaceutical and food industries.

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This study aimed to determine the bioactive compounds of Bacillus sp. MERNA97 extract and to evaluate their efficacy against the oxidative damage, genotoxicity, chromosomal aberration and DNA fragmentation in rats treated with AFB1. Sixty male Sprague-Dawley rats were divided into 6 groups and treated for 6 weeks and included the control group, AFB1-treated group (80 µg/kg b. w), the groups treated with Bacillus extract (BE) at low (2 mg/kg b.w) or high (4 mg/kg b.w) dose and the groups treated with AFB1 plus BE at the two doses. Blood and tissues samples were collected for different assays. The GC-MS results revealed the isolation of 44 compounds belong to different classes. The in vivo results showed that AFB1 disturbs all the biochemical parameters, oxidative stress markers, cytokines gene expression chromosomal aberration and DNA fragmentation along with the histological changes in the liver tissue. BE at the two tested doses induced a significant improvement in all parameters tested and the histological picture in a dose dependent manner. It could be concluded that the extract of Bacillus sp. MERNA97 isolated from the marine environment in the Red Sea is a promise as a source of novel compounds with therapeutically benefits.

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Heavy metals are considered a main public health hazards, they are known to accumulate in fruits, which are heavily consumed by humans because of their unique sweet taste and potential health benefits. Therefore, the aim of this study was to determine the possible contamination levels of cadmium (Cd), chromium (Cr), copper (Cu), lead (Pb) and nickel (Ni) in some selected fruits obtained from four Egyptian governorates and to compare the contamination levels with those of the recommended permissible limits. Results revealed that Pb and Cd were absent in all fruit samples, while Cr was only detected in grapes obtained from Cairo and Fayoum governorates and exceeded the maximum permissible limit (0.10 mg kg-1). Nickel and Cu were detected in all fruit samples. Nickel was reduced more than copper after washing process of grapes especially in samples obtained from Alexandria and Giza governorates. After peeling process, Cu was extremely reduced in orange samples obtained from the following governorates in descending order Cairo, Alexandria, El-Fayoum and Giza. Estimated daily intake of heavy metals in fruit samples were found to be higher than that of the tolerable daily intake, indicating potential risk to human health. Therefore, to decrease the risk to human health, fruits must be washed well before eating to decrease heavy metal concentrations.

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This study aimed to assess the effect of encapsulating humic acid inside yeast cell walls (YCW) to detoxify AFB1 in in vitro gastrointestinal models. Glucan Mannan Lipid Particles (GMLPs) from Saccharomyces cerevisiae cell walls showed the highest AFB1 adsorption in simulated gastric fluid (SGF) after 10 min, and in simulated intestinal fluid (SIF) after 1 h. GMLPs are hollow 3-4 micron porous microspheres that provide an efficient system for the synthesis and encapsulation of AFB1-absorbing nanoparticles (NPs). Humic acid nanoparticles (HA-NPs) were synthesized within the GMLP cavity by complexation with ferric chloride. Encapsulating HA-NPs in GMLPs increased HA-NP stability in SIF. The hybrid GMLP HA-NP formulation synergistically enhanced AFB1 binding compared to individual GMLP and HA components in SGF and in SIF. Cytotoxicity on a murine macrophage cell line demonstrated that GMLP HA-NP-AFB1 complexes were stable in both SGF and SIF, detoxified AFB1 and are suitable for in vivo testing.

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Agarwood (Oudh), is often used by people in the Kingdom of Saudi Arabia. The Oudh has been mentioned in the Hadith and is traditionally used for its aroma (perfuming smell) and potential medicinal applications. The aim of the study was to isolate mycotoxigenic fungi that grow on agarwood and the factors and storage conditions that enhance their growth potential. In addition to the detection of associated mycotoxins like: Aflatoxin B1 (AFB1) and ochratoxin A (OTA) from agarwood. Agarwood samples were collected from local markets of Jeddah governorate, Kingdom of Saudi Arabia. Standard dilution plate method was used for the isolation of fungi. Isolated fungi were identified based on morphological characteristics and confirmed using molecular biology techniques. AFB1 and OTA were detected by High Performance Liquid Chromatography (HLPC). The results indicated that the most commonly isolated fungal genera were in the following descending order: Aspergillus, Penicillium, Fusarium and Rhizopus. Among Aspergillus genera, A. flavus and A. ochraceus were detected based on their morphology and confirmed by PCR using specific primers. It was also noted that AFB1 was released by 15.3 and 55.0% of A. flavus and A. parasiticus isolates respectively with levels reaching up to 14.60 µg/L. The moisture content in the samples ranged from 3% to 10% affected fungal growth. AFB1 was detected in 22 out of 50 of the samples. The maximum level of AFB1 (50.7 µg/kg) was detected in samples with higher moisture content (12%) stored at a temperature of 32 °C. Aspergillus fungi were found to be the most predominant fungal genera found on agarwood. Moisture content (9-10%) and storage temperature (32 °C) stimulated fungal growth and their ability to produce mycotoxins. For this reason, storage conditions at the marketing place should be adequate in order not to provide a conducive environment for fungal growth which is associated with the mycotoxin production. In order to prevent fungal growth and mycotoxin production, it would be recommended to store agarwood at temperatures not exceeding 25 °C and moisture content of up to a maximum of 5-6%.

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PURPOSE: Zearalenone is a potent estrogenic metabolite produced by some Fusarium and Gibberella species. Therefore, the aim of this study was to determine the accumulation of Technetium-[99m]-labeled zearalenone in mice organs as well as to study the protective effect of Lactobacillus plantarum. MATERIALS AND METHODS: Two different groups of mice were used; the first group of mice received Technetium-[99m]-zearalenone only, and the second group received both Technetium-[99m] zearalenone and L. plantarum (as a protective agent). Technetium-[99m]-labeled zearalenone was administered to mice by tail vein injection and L. plantarum was administered orally. Accumulation of labeled zearalenone in mice was monitored for 120 min. RESULTS: Labeled zearalenone was distributed primarily in the intestine (44.5%) and liver (20.12%) after 120 min in the first group of mice. In the second group of mice which received both Technetium-[99m] zearalenone and L. plantarum, labeled zearalenone was accumulated primarily in the intestine (46.8%) and liver (18.9%). These results indicated the decrease in labeled zearalenone in the liver due to the effect of lactic acid bacteria. CONCLUSION: Technetium-[99m]-labeled zearalenone can be used as a new tracer for organ imaging and that L. plantarum can decrease the bioavailability of zearalenone in mice organs.

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This study aimed to determine the bioactive compounds of actinomyces (ACT) isolated from the Egyptian environment (D-EGY) and to evaluate their protective activity against AFB1 in female Sprague-Dawley rats. Six groups of animals were treated orally for 3 weeks included: C, the control group, T1, AFB1-treated group (80 µg/kg b.w), T2 and T3, the groups received ACT extract at low (25 mg/kg b.w) or high (50 mg/kg b.w) doses, T4 and T5, the groups received AFB1 plus the low or high dose of ACT extract. Blood, bone marrow and tissue samples were collected for different analyses and histological examination. The results revealed the identification of 40 components, representing 99.98%. Treatment with AFB1 disturbs liver function parameters, oxidative stress markers, antioxidant gene expressions, DNA fragmentation and induced severe histological changes. ACT extract at the low or high doses did not induce significant changes in all the tested parameters or histological picture of the liver. Moreover, ACT extract succeeded to induce a significant protection against the toxicity of AFB1. It could be concluded that the bioactive compounds in ACT are promise candidate for the development of food additive or drugs for the protection and treatment of liver disorders in the endemic area.

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Lactic acid bacteria (LAB) have been reported to remove mycotoxins from aqueous solutions through a binding process, which appears to be species and strain specific. The aim of the current study was to evaluate the protective role of Lactobacillus casei (L1) and Lactobacillus reuteri (L2) against aflatoxin (AFs)-induced oxidative stress in rats. Sixty female Sprague-Dawley rats were divided into 6 groups including the control group and the groups treated with L1 or L2 (1 x 10¹¹/ml) alone at a dose of 10 ml/kg b.w or plus AFs (3 mg/kg diet) for 4 weeks. At the end of the treatments, blood and tissue samples were collected for biochemical and histological studies. The results indicated that AFs alone induced a significant decrease in food intake and body weight and a significant increase in transaminase, alkaline phosphatase cholesterol, triglycerides, total lipids, creatinine, uric acid and nitric oxide in serum and lipid peroxidation in liver and kidney accompanied with a significant decrease in total antioxidant capacity. Treatments with L1 or L2 succeeded to induce a significant improvement in all the biochemical parameters and histological picture of the liver. Moreover, L2 was more effective than L1 and both can be used safely in functional foods.

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The leafy parts of thyme and its essential oil have been used in foods for the flavor, aroma and preservation and also in folk medicines. The aim of the current study was to determine the components of Thymus vulgaris L essential oil and to evaluate the protective effects of this oil against aflatoxin-induce oxidative stress in rats. Thirty six mature male Sprague-Dawley were divided into six treatment groups and treated for 2 weeks as follows: control group; the groups treated orally with low and high doses of T. vulgaris oil (5 and 7.5 mg/kg b.w.); the group fed AFs-contaminated diet (2.5 mg/kg diet) and the groups fed AFs-contaminated diet and treated orally with the oil at the two tested doses. Blood and tissue samples were collected at the end of treatment period for biochemical study and histological examination. The results indicated that the oil contains Carvarcrol (45 mg/g), Thymol (24.7 mg/g), ß-Phellandrene (9.7 mg/g), Linalool (4.1 mg/g), Humuline (3.1 mg/g), α-Phellandrene (2.3 mg/g) and Myrcene (2.1 mg/g). However, α and ß-pinene, Myrcene, α-thyjone, Tricyclene, 1, 8-cineole, and ß-sabinene were found in lower concentrations. Treatment with AFs alone disturbs lipid profile in serum, decreases Total antioxidant capacity, increase creatinine, uric acid and nitric oxide in serum and lipid peroxidation in liver and kidney accompanied with a sever histological changes in the liver tissues. The oil alone at the two tested doses did not induce any significant changes in the biochemical parameters or the histological picture. The combined treatment showed significant improvements in all tested parameters and histological pictures in the liver tissues. Moreover, this improvement was more pronounced in the group received the high dose of the oil. It could be concluded that the essential oil of T. vulgaris has a potential antioxidant activity and a protective effect against AFs toxicity and this protection was dose dependent.

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BACKGROUND: Aflatoxins are a group of mycotoxins that cause serious chronic disease outbreaks and contaminate several food products such as corn and its by-product, corn gluten. The aim of the current study was to evaluate the effect of hydrochloric acid (HCl) on aflatoxin B(1) (AFB(1) ) degradation in contaminated corn gluten under different HCl concentrations, hydrolysis temperatures and hydrolysis times. RESULTS: During the wet milling process the highest AFB(1) level (45.68 µg kg(-1) ) (37.86%) was found in corn gluten fraction. Treatment with 1 mol L(-1) HCL at 110 °C resulted in degradation of AFB(1) by 27.6% (33.07 µg kg(-1) ) after 4 h and reached 42.5% (26.26 µg kg(-1) ) after 8 h. Increasing HCl concentration from 1 to 3 mol L(-1) HCl resulted in increased degradation of AFB(1) , while complete degradation occurred in the presence of 5 mol L(-1) HCl after 4 h at 110 °C. Meanwhile, half-life time of AFB(1) was recorded after 2 h at 100 °C and was < 2 h at 110 °C in the presence of 3 mol L(-1) HCl. CONCLUSION: It could be demonstrated that the manufacture of hydrolyzed vegetable protein is a suitable method for decontamination of aflatoxin in highly contaminated grains, especially gluten fractions. The hydrolysis reaction could be considered in terms of first-order reaction kinetics of AFB(1) degradation.

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