Isolation and characterization of a Bacillus subtilis strain with aflatoxin B1 biodegradation capability

Aflatoxins are type of mycotoxins mainly produced by Aspergillus flavus and a common contaminant of food and grain, posing a serious economic and health problem worldwide. In order to find efficient bacteria to remove or detoxify these mycotoxins, a bacterial strain capable of degrading aflatoxin B₁ (AFB₁) was isolated from soil samples using a culture medium containing coumarin as the sole carbon source. Based on 16S rRNA gene sequence analysis, this isolate was identified as Bacillus subtilis JSW-1; its further characterization showed that it could inhibit the growth of A. flavus with an inhibition ratio of 58.3% and could degrade AFB₁ by 67.2% after incubation at 30 °C for 72 h. The aflatoxin B₁-degrading activity of isolate JSW-1 was predominantly attributed to the cell-free supernatant and this activity was found to be heat stable but sensitive to proteinase K treatment, indicating that the extracellular proteins or enzymes are responsible for the AFB₁ degradation. In addition, no degradation products of AFB₁ could be detected by liquid chromatography-mass spectrometry (LC-MS) analysis, indicating that the parent AFB₁ might be biotransformed to compounds with chemical properties different from that of AFB₁.     

Detection of aflatoxin B1 by aptamer-based biosensor using PAMAM dendrimers as immobilization platform

We report an aptamer-based biosensor for detection of aflatoxin B₁ (AFB₁), a mycotoxin identified as contaminant in food. The sensor is assembled in a multilayer framework that utilizes cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) for acquiring the signal response by means of redox indicators: K[Fe(CN)₆]^−3/−4. Poly (amidoamine) dendrimers of fourth generation (PAMAM G4) immobilized on gold electrode covered by cystamine, were employed for attachment of single stranded amino-modified DNA aptamers specific to AFB₁. The cystamine-dendrimers (Cys-PAMAM) layers were compared with other immobilization platforms such as cystamine (Cys), 11-mercaptoundecanoic acid (MUA) and 11-mercaptoundecanoic acid-dendrimers (MUA-PAMAM), being the first approach the most appropriate for producing sensitive and reproducible signal in the range of concentrations 0.1–10 nM AFB₁. The sensor was validated in certified contaminated peanuts extract as well as in spiked samples of peanuts-corn snacks and the sensing response was evaluated and compared in terms of the matrix effect. The aptamer specificity was analyzed by testing the sensor in other mycotoxins such as aflatoxin B₂ (AFB₂) and ochratoxin A (OTA). The limit of detection achieved by this sensor was LOD = 0.40 ± 0.03 nM, it was regenerable in 0.2 M glycine-HCl and it did not lose its stability up to 60 h storing at 4 °C. Atomic Force Microscopy (AFM) studies were also performed for illustrating individual steps of biosensor assembly.     

Incidence and consumer awareness of toxigenic Aspergillus section Flavi and aflatoxin B1 in peanut cake from Nigeria

Peanut cake samples were collected from major markets in five states of Nigeria and evaluated for incidence of toxigenic Aspergillus section Flavi populations, and aflatoxin B₁ (AFB₁) levels by liquid chromatography tandem mass spectrometry (LC–MS/MS). The awareness of consumers to the presence of aflatoxin in the snack and potential health risks of its regular ingestion was evaluated by questionnaire analysis. Aspergillus section Flavi populations were recovered from 83% of the peanut cake samples. Aspergillus flavus L-strain was the most predominant (>56%) across the states while Aspergillus tamarii had the least mean incidence (2.7%). The incidence of atoxigenic strains was significantly (p < 0.05) higher than that of toxigenic strains in samples from Lagos and Kaduna, while the toxigenic strains had a significantly (p < 0.05) higher incidence than the atoxigenic strains in Niger. All analyzed cake samples contained AFB₁ in concentrations exceeding the NAFDAC recommended level for AFB₁ in food and reaching up to 2824 μg/kg. There was a weak positive correlation (r = 0.32, p = 0.03) for the relationship between the incidence of toxigenic strains in the samples and AFB₁ concentration. The consumer awareness data showed that 64% of the respondents consumed peanut cake; majority of who are youth of economic and reproductive age. Eighty-five percent of the consumers lacked awareness of aflatoxin contamination in the snack and possible health risks associated with its ingestion.     

Removal of aflatoxin B1 by roasting with lemon juice and/or citric acid in contaminated pistachio nuts

Nowadays, aflatoxin B₁ (AFB₁) could be considered as one of the most hazardous mycotoxins for humans, and nuts comprise one of the major responsible food categories for human exposure to this mycotoxin. Thus, complete elimination of AFB₁ or reduction of its content in nut foods, such as pistachio attracted lots of attentions. In the current study, the efficacy of roasting process by incorporation of lemon juice and/or citric acid on the reduction of AFB₁ in contaminated pistachio nuts (AFB₁ at two levels of 268 and 383 ng/g) was investigated. Significant degradation of AFB₁ (up 93.1% for AFB₁) was recorded by applied treatment protocols. Although roasting of 50 g pistachio nuts with 30 ml water, 30 ml lemon juice and 6 g of citric acid at 120 °C for 1 h resulted to a significant degradation (93.1 ± 8.2%) of AFB₁, this treatment altered the desired physical properties. Roasting with 30 ml water, 15 ml lemon juice and 2.25 g of citric acid at 120 °C for 1 h reduced the level of AFB₁ in 49.2 ± 3.5% of the initial level without a noticeable change in desired appearance of pistachios. Hence, a synergistic effect between heating and lemon juice/citric acid in order to AFB₁ degradation was observed. It could be concluded that roasting process with lemon juice and citric acid could be applied as a useful and safe degradation method of AFB₁ in naturally contaminated pistachio nuts.     

Effects of high pressure processing on immunoreactivity and microbiological safety of crushed peanuts

Peanuts are a common economical food source consumed worldwide but exist health concern of food allergy and are particularly susceptible to infection by the mold fungus Aspergillus flavus during storage, accumulating highly toxic substance aflatoxin. In this study, the effect of high pressure treatments on peanut immunoreactivity, peanut amino acid composition, A. flavus growth and aflatoxin contents on crushed peanuts was evaluated. Results showed that immunoreactivity of peanuts treated with 600 MPa and 800 MPa for 10 min was significantly lower (P < 0.05) than those of the control group by 69.2 ± 5.3% and 73.3 ± 1.9%, respectively. High pressure treatment at 800 MPa decreased total essential amino acid content as well as two nutritional indexes, the chemical score and the essential amino acid index, by 32.4 ± 2.1% and 31.1 ± 3.2%, respectively. The growth of aflatoxigenic fungi was inhibited in peanuts with aflatoxin accumulation that were subjected to different levels of pressure treatments during 30 days of storage. Peanuts treated with 600 MPa and 800 MPa had considerably lower aflatoxin levels, 0.26 μg/g and 0.22 μg/g in wet basis, respectively, than the control peanut aflatoxin level (9.08 μg/g) on day 30. Results were demonstrated that high pressure treatment had a significant inhibitory effect on A. flavus growth in peanuts and this contributes to reduction of aflatoxin production and accumulation instead of directly destroy aflatoxin. Taken together, the findings of this study indicated that high pressure treatment could preserve peanut quality by reducing food immunoreactivity and by eliminating A. flavus in peanuts.     

The in vitro ability of different Saccharomyces cerevisiae – Based products to bind aflatoxin B1

The purpose of this study was to evaluate the ability of Saccharomyces cerevisiae (SC) – based products from sugar cane fermentation (dried yeast – DY, autolyzed yeast – AY, cell wall – CW) and from beer fermentation (brewery dehydrated residue – BDR) to bind aflatoxin B₁ (AFB₁) in phosphate buffer saline (PBS) spiked with 0.5 μg AFB₁ mL⁻¹. All SC cells were heat-killed (121 ° C, 10 min) and then used for checking the effect of contact time (5, 10, 20 and 30 min) on toxin binding capacity. Compared to the CW and BDR treatments, DY and AY had higher (p < 0.05) capability to bind AFB₁ in PBS, although there were no differences (p > 0.05) among the contact times for any product evaluated. The mean percentages of AFB₁ bound by the DY were higher than AY, varying from 96.5% to 99.3% and from 90.4% to 97.5%, respectively, although the differences were not significant (p > 0.05). The SC-based products from sugar cane fermentation have a potential application for reducing levels of AFB₁ in contaminated food products. However, additional studies are needed to investigate the mechanisms involved in the removal process of toxin by SC and factors that affect toxin sequestration aiming the commercial application in food industry.     

Antifungal, aflatoxin inhibition and antioxidant activity of Callistemon lanceolatus (Sm.) Sweet essential oil and its major component 1,8-cineole against fungal isolates from chickpea seeds

The present investigation reports the extent of molds and aflatoxin contamination to Avarodhi, Kabuli, Pusa 256, Radha and Samrat varieties of chickpea seeds. The study also examines the chemical composition of Callistemon lanceolatus (Sm.) Sweet essential oil and its antifungal, antiaflatoxin and antioxidant activity. During standardization of chemical profile, a total of 8 compounds constituting 0.862 mg/mL of oil composition were analyzed by GC-MS analysis where 1,8-cineole was recorded as a major component (0.56 mg/mL). The antifungal activity of EO and 1,8-cineole was evaluated by contact assay on Czapek’s dox agar. The EO (0.227-0.908 mg/mL) and 1,8-cineole (0.918 mg/mL) showed remarkable antifungal effect against all the fungal isolates of chickpea. Their minimal inhibitory (MIC) and fungicidal (MFC) concentrations for Aspergillus flavus were lower than those of the prevalent systemic fungicide Nystatin. Aflatoxin B₁ (AFB₁) production by NKD-208 isolates of A. flavus was strongly inhibited even at the lower fungistatic concentration of EO and 1,8-cineole.There was no adverse effect of EO treatment on chickpea seed germination suggesting its non-phytotoxic nature. Based on the findings of present investigation, C. lanceolatus essential oil may be recommended as botanical preservative for the enhancement of shelf life of food items in- view of the adverse effect of synthetic preservatives and its strong antifungal, aflatoxin inhibition and antioxidant activity.     

Investigations on the essential oil of Lippia rugosa from Cameroon for its potential use as antifungal agent against Aspergillus flavus Link ex. Fries

Lippia rugosa essential oil was tested for its effectiveness against Aspergillus flavus on artificial growth media. The chemical composition of the oil was determined by gas chromatography–mass spectrometry (GC–MS). Geraniol (51.5%), nerol (18.6%) and geranial (10.4%) were the main components of Lippia oil. After 8 days of incubation on essential oil supplemented medium, mycelium growth of A. flavus was totally inhibited by 1000 mg l^?1 of L. rugosa essential oil. The effect of essential oil on aflatoxin B₁ synthesis was evaluated in SMKY broth. The medium supplemented with different essential oil concentrations, was inoculated with A. flavus mycelium and incubated at 25 °C. After 2, 4, 6 and 8 days, aflatoxin B₁ (AFB₁) was quantified in the supernatant using Enzyme Linked Immuno-Sorbent Assay (ELISA). Results showed that aflatoxin B₁ synthesis was inhibited by 1000 mg l^?1 of L. rugosa essential oil after 8 days of incubation. The effect of the EO on the H⁺-ATPase pumping membrane was also evaluated in the presence of several concentrations of oil (200–2000 mg l^?1) by monitoring glucose-induced acidification of the external medium. L. rugosa essential oil at the concentration of 2000 mg l^?1 completely inhibited the activity of this enzyme. These data suggest that the essential oil of L. rugosa is a fungicidal for A. flavus and its possible cellular target include the H⁺-ATPase.Results obtained in the present study indicate the possibility of exploiting Lippia rugosa essential oil in the fight against strains of A. flavus responsible for biodeterioration of stored foods products.     

Development and validation of an accurate and rapid LC-ESI-MS/MS method for the simultaneous quantification of aflatoxin B1, B2, G1 and G2 in lotus seeds

An accurate and rapid LC-ESI-MS/MS analytical method was developed and validated for the simultaneous determination of aflatoxin B₁, B₂, G₁ and G₂ in lotus seeds. The samples were firstly extracted with methanol-water solution (80:20, v/v), and then cleaned up by immunoaffinity columns. The mass spectrometer was operated in the positive ionization electrospray (ESI+) mode using multiple reaction monitoring (MRM) for analysis of four aflatoxins. The transitions of m/z 313 → 285 (AFB₁, CE 33 eV), m/z 315 → 259 (AFB₂, CE 37 eV), m/z 329 → 243 (AFG₁, CE 37 eV) and m/z 331 → 257 (AFG₂, CE 37 eV) were used to quantify these four aflatoxins, respectively. The limits of detection (LODs) of aflatoxin B₁, B₂, G₁ and G₂ were 0.007, 0.005, 0.003 and 0.005 μg kg^?1 based on a signal-to-noise ratio of 3:1, respectively. The limits of quantification (LOQs) of aflatoxin B₁, B₂, G₁ and G₂ were 0.02, 0.015, 0.01 and 0.015 μg kg^?1 based on a signal-to-noise ratio of 10:1, respectively. Recoveries for samples of spiked lotus seeds were all above 66% with relative standard deviation all below 15% for all compounds. Nineteen out of twenty batches of lotus seeds collected from different drug stores or markets in China were found to be contaminated with aflatoxins at different levels ranging from 0.02 to 688.4 μg kg^?1.     

Effectiveness of pulsed light treatment for degradation and detoxification of aflatoxin B1 and B2 in rough rice and rice bran

Aflatoxins primarily accumulate in the hull and bran layers of rough rice making these by-products of rice milling unsuitable for animal feed or human consumption. Contaminated rough rice is also a potential source of aflatoxin exposure to workers handling the grain during post-harvest storage and processing. Currently, no technologies are available to remove or detoxify these toxic and mutagenic fungal metabolites from contaminated rough rice. Pulsed light (PL) is a novel technology with the potential to degrade and detoxify aflatoxins in foods and their processing by-products. Rough rice was inoculated with Aspergillus flavus to produce aflatoxin B₁ (AFB₁) and B₂ (AFB₂) contamination, followed by PL treatments of 0.52 J/cm²/pulse for various durations. A PL treatment time of 80 s reduced AFB₁ and AFB₂ in rough rice by 75.0% and 39.2%, respectively; while a treatment time of 15 s reduced AFB₁ and AFB₂ in rice bran by 90.3% and 86.7%, respectively. Since PL treatments result in the degradation of aflatoxins in situ, the toxicity and mutagenic activity of the residual by-products of AFB₁ and AFB₂ after PL treatment were evaluated. Toxicity was estimated using the brine shrimp (Artemia salina) lethality assay and mutagenicity measured by the fluctuation test with Salmonella typhimurum tester strains TA98 and TA100. The mutagenic activity of AFB₁ and AFB₂ was completely eliminated by PL treatment, while the toxicity of these two aflatoxins was significantly decreased. The obtained results suggest that PL technology has a promising potential to degrade, detoxify, and inactivate the mutagenic activity of aflatoxins in rough rice and rice bran.     

Annual and regional variations of aflatoxin B1 levels seen in grains and feed coming from Croatian dairy farms over a 5-year period

The aim of the study was to investigate annual and regional differences in the level of aflatoxin B₁ (AFB₁) in grains and dairy cattle feed. Maize (n = 972), wheat (n = 201), barley (n = 147), oat (n = 136), grain mixtures (n = 168), and dairy cattle feed (n = 325) were sampled from 2009 to 2013 on different farms and in different farm factories situated in four Croatian regions. The samples were analysed for AFB₁ using the validated ELISA immunoassay. AFB₁ was determined in 16.4% of all investigated samples, among which maize was proven to be the most contaminated, with 21.7% of the samples recovered during 2013 harbouring AFB₁ in concentrations over the permissible ones. Levels higher than permitted were observed in 17.9% and 12.3% of grain mixtures and dairy cattle feed, respectively, whereas concentrations of AFB₁ determined in other crops throughout the investigated period met the stipulated requirements. The results revealed the AFB₁ occurrence to be significantly (p < 0.05) dependent on the cultivation region, with the highest levels generally found in maize harvested in 2013 and consequently in grain mixtures and cattle feed that can most likely be associated with climatic conditions as the most critical factor for mould formation, and thus also AFB₁ production.     

Inhibition of non-toxigenic Aspergillus niger FS10 isolated from Chinese fermented soybean on growth and aflatoxin B1 production by Aspergillus flavus

A non-toxigenic strain FS10 was isolated from Chinese fermented soybean to control AFB₁ contamination and conformed as Aspergillus niger by 18S rDNA. To elucidate the biocontrol mechanism, the ability of culture filtrate of strain FS10 against growth and AFB₁ production by toxigenic Aspergillus flavus was evaluated in vitro. Meanwhile, the influence of filtrate on spore morphology of A. flavus was analyzed by optical microscope and atomic force microscope (AFM). Results demonstrated that the culture filtrate of FS10 significantly suppressed A. flavus growth, sporulation and AFB₁ biosynthesis. After 9 days, the inhibition of mycelia growth, sporulation and AFB₁ production reached to 37.7%, 100% and 94.5%, respectively. In addition, optical microscopy and AFM showed that the culture filtrate serious damaged spore morphology, accompanying with spore analysis and cell wall collapse, resulted in inhibition of spore germination. The degradation activity of FS10 culture filtrate on AFB₁ was also estimated. When AFB₁ was incubated with the culture filtrate, 85.8% of AFB₁ could be degraded after 5 days. It is concluded that the culture filtrate of strain FS10 has pleiotropic effects on control AFB₁ contamination. As a safe, effective and economic biological agent, the filtrate could be applied in fermented food to prevent toxigenic Aspergillus spp. and AFB₁ contamination.     

Inactivation of aflatoxigenic fungi and the reduction of aflatoxin B1 in vitro and in situ using gamma irradiation

Detailed investigation on the effect of gamma (γ) irradiation on germination, sporulation, and growth of aflatoxigenic moulds (Aspergillus parasiticus 2999, Aspergillus flavus 305, and Aspergillus niger 388), as well as on the reduction of aflatoxin B₁ (AFB₁) level in artificially and naturally contaminated maize/feed samples was performed. The results of in vitro and in situ experiments with aflatoxigenic moulds demonstrated that 5 kGy-γ irradiation manages to prevent sporulation, germination and growth of the tested moulds both when in form of a pure and when in form of a mixed culture. In the feed samples artificially contaminated with AFB₁ (50 μg kg⁻¹) 5 kGy-γ irradiation reduced AFB₁ level by around 60%, while 10 kGy-dose reduce it for around 85%. Similarly, in feed samples spiked with AFB₁ in the concentrations of 100 μg kg⁻¹ 5 kGy-dose reduced the AFB₁ level by approximately 70%, while the dose of 10 kGy reduced it by approximately 90%. The experiments on naturally contaminated maize samples (n = 30) confirmed these observations; following a 5 kGy-irradiation, the overall mean AFB₁ reduction equalled to 69.8%, while the irradiation with a 10 kGy-dose achieved the overall mean toxin reduction of 94.5%. The obtained results indicate that γ irradiation can be used to prevent the growth of aflatoxigenic moulds and to reduce the AFB₁ levels in various goods intended for animal and human consumption, thus minimizing the animal and human exposure to this carcinogenic mycotoxin.     

Effect of Hibiscus sabdariffa extract and Nigella sativa oil on the growth and aflatoxin B1 production of Aspergillus flavus and Aspergillus parasiticus strains

This study was undertaken to evaluate the inhibitory effect of Hibiscus sabdariffa calyx extract at concentrations of 5, 7.5, 10 and 12.5 g/100 ml and Nigella sativa oil at concentrations of 1, 2 and 3 ml/100 ml on the growth and aflatoxin B₁ production by Aspergillus parasiticus (CBS 921.7) and Aspergillus flavus (SQU 21) strains. The inhibition of aflatoxin B₁ production by the different concentrations of H. sabdariffa calyx ranged between 91.5-97.9% and 87.1-93.3% for A. flavus and A. parasiticus strains, respectively, whereas the inhibition by different concentrations of N. sativa oil ranged between 47.9 and 58.3% for A. flavus and 32-48% for A. parasiticus strains. The different concentrations of H. sabdariffa calyx and N. sativa oil had no significant effect on the growth of either Aspergillus species. Neither H. sabdariffa calyx nor N. sativa oil detoxified pure aqueous aflatoxin. Our results suggest that H. sabdariffa calyx and N. sativa oil extracted from seeds had metabolic effects on aflatoxin biosynthesis pathway of both Aspergillus species and can be used as an effective biocontrol and non-toxic biopreservatives in food industry against aflatoxin contamination.     

Mycoflora and absence of aflatoxin contamination of commercialized cassava chips in Benin, West Africa

Studies conducted in Benin, in which the main staple foods are maize, cassava, groundnuts and yams, showed high levels of aflatoxin residues in blood of the exposed population. The natural contamination with fungi and aflatoxins in cassava chips sold at markets in Benin, West Africa was investigated. A total of sixty samples were sampled from open markets in 11 districts of 3 agroecological zones and analyzed for the presence of mycoflora and aflatoxin B₁, B₂, G₁ and G₂. Fourteen genera of fungi were associated with marketed dried cassava chips. Within these, twenty- two isolates were identified to species level, whereas four were identified only to genus. The dominating fungal species isolated were Rhizopus oryzae, Nigrospora oryzae, Chrysonilia sitophila, Cladosporium resinae, Cladosporium herbarum, Apergillus niger and Aspergillus flavus. Fifty-four out of sixty samples were contaminated with A. flavus. The rate of occurrence in CFU/g of A. flavus fungi was lower than for all other fungal species together. Aflatoxin was not detected in any of the samples analyzed using HPLC with post-column photochemical derivatization and fluorescence detection. The limit of detection (LOD) was 0.1 μg/kg. Results from this study suggest cassava chips are unlikely to be a source of aflatoxin in Benin, and that other staples such as maize and groundnuts are more important in aflatoxin exposure. Therefore it can be speculated that staples like maize and groundnut are more important in aflatoxin exposure.     

Impact of three sterol-biosynthesis inhibitors on growth of Fusarium langsethiae and on T-2 and HT-2 toxin production in oat grain under different ecological conditions

The health-beneficial properties of oats have led to an increase in the consumption of oats and oat-based food products in recent years. Fusarium langsethiae grows on small grain cereals, especially oats and can result in contamination with type A trichothecenes (T-2 toxin (T-2) and HT-2 toxin (HT-2)) in crops pre- and post-harvest. The aim of this work was to assess the efficacy of three fungicides (fenpropimorph, prochloraz and tebuconazole) on temporal growth of two F. langsethiae strains and the accumulation of T-2 and HT-2 in oat grains, under different water activities (a_w; 0.95 and 0.98) and temperatures (15 and 25 °C). All the antifungal agents reduced growth rates when compared to controls, and this increased with increasing fungicide dose. The ranges of ED₅₀ values (mg/kg) were 65–270 for fenpropimorph, 0.25–4.2 for prochloraz, and 0.3–14 for tebuconazole. The ED₉₀ values (mg/kg) ranged from 170 to > 800, 0.5 to > 10 and 0.5 to > 15 for these three fungicides respectively. The ED₅₀ values were not statistically significantly affected by the factors a_w, temperature or strain. However, there were significant differences among the fungicides. Fenpropimorph proved less efficient that the two azoles, which do not differ significantly. In general, levels of HT-2 were higher than T-2 in all cultures regardless of environmental conditions. Overall, HT-2 concentration was always higher at 25 than 15 °C and increased from day 14 to day 21. Levels of both toxins generally decreased with increasing fungicide dose regardless of fungicide type, strain and incubation time. No toxins were detected in cultures at 0.95 a_w in the presence of any of the three fungicides. Also under wetter conditions at 0.98 a_w neither mycotoxin was found in cultures treated with prochloraz at doses >1 mg/kg (15 °C) or >3 mg/kg (25 °C) or tebuconazole at doses >6 mg/kg (15 or 25 °C). ANOVA showed that in treatments with each fungicide the factors dose, time and temperature significantly affected toxin production while there was no difference between the strains.     

Effect of ozone treatment on aflatoxin B1 and safety evaluation of ozonized corn

This paper studies the ozone treatment effect on degradation of aflatoxin B₁ (AFB₁) in corn with different moisture content (MC). The toxicity of the degradation products (DPs) of the ozone-treated AFB₁-Contaminated Corn (ACC) was also evaluated using the human hepatocellular carcinoma cell line (HepG2) as model cells. The degradation rate of AFB₁ in corn increases with ozone concentration and treatment time. The results showed that ACC with 13.47% MC was easier to be degraded by ozone than with 20.37% MC. Treated with 90 mg L⁻¹ ozone for 20 min and 40 min, AFB₁ in corn with 13.47% MC decreased from 83 μg kg⁻¹ to 18.12 μg kg⁻¹ and 9.9 μg kg⁻¹, respectively, well meeting the China National Standard of AFB₁ in corn (20 μg kg⁻¹). In order to evaluate the safety of ozone used on ACC, the impacts of AFB₁ as well as untreated and ozone-treated ACC with the same level of AFB₁ content on HepG2's survival rate, morphology, and apoptosis were studied. The results showed that ACC had high cell toxicity while the toxicity of ozone-treated ACC had no significant difference with that of the AFB₁-free culture solution. It is concluded that ozonation can quickly and effectively degrade AFB₁ in corn and diminish ACC's toxicity, and therefore, ozonation is expected to be an effective, fast, and safe method for AFB₁ degradation in ACC.     

Aflatoxin B1 in post-harvest peanuts and dietary risk in China

To monitor the aflatoxin contamination status in raw peanuts and evaluate the effect on public health, 1040 samples were collected from four agro-ecological zones throughout 12 provinces from 2009 to 2010 in China and then analyzed for aflatoxin B₁ (AFB₁) levels using High Pressure Liquid Chromatography (HPLC) and immunoaffinity columns. The results revealed that AFB₁ was detected in 25% of the samples, ranging from 0.01 to 720 μg/kg. The Monte Carlo and bootstrap methods were employed to estimate AFB₁ intake in children and adults and their potential liver cancer risk. The mean estimated intakes for children and adults were 0.218-0.222 ng/kg body weight (bw)/day and 0.106-0.108 ng/kg bw/day. The liver cancer risk, calculated by two approaches derived from the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and European Food Safety Authority (EFSA), were estimated at 0.003-0.17 cancer cases/year/100,000 and 24.7-1273 margins of exposure values, respectively. The results suggest that AFB₁ contamination in raw peanuts and dietary risk was low, but essential surveillance measures should be taken to protect public health.     

Natural occurrence of aflatoxin B1, ochratoxin A and citrinin in Croatian fermented meat products

When domestic animals are exposed to mycotoxins, significant amounts of the latter shall be carried over into animal products such as milk, eggs and meat. This study was carried out in order to determine the possible presence of aflatoxin B₁ (AFB₁), ochratoxin A (OTA) and citrinin (CIT) in game sausages (n = 15), semi-dry sausages (n = 25) and fermented dry-meat products (n = 50), randomly taken from individual producers and the Croatian market. AFB₁ and OTA were quantified using ELISA, while CIT was quantified using HPLC-fluorescence detector. Out of 90 samples, the fungi most frequently isolated from dry-cured meat products were of Penicillium species, while Aspergillus was isolated from only one sample. As much as 68.88% of the samples were positive for mycotoxins. Finally, the analysis of different types of meat products resulted in OTA identification in 64.44%, CIT identification in 4.44% and AFB₁ identification in 10% of the samples. The maximum OTA concentrations established in the commercial sausage samples equalled to 7.83 μg/kg, while that of AFB₁ amounted to 3.0 μg/kg. Generally, although OTA was detected in all three types of products in different percentage shares, mutual differences were not statistically significant (P > 0.05).