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.     

Biological detoxification of Aflatoxin B1 by Bacillus licheniformis CFR1

Aflatoxin B₁ is the most harmful among the mycotoxins commonly present in food and feed, and it may lead to hepatocellular carcinoma in humans and animals. Therefore, limiting its exposure to humans and livestock is very much essential. The present study aims to isolate and characterize Aflatoxin B₁ detoxifying bacteria from various sources, to develop a safe and environment-friendly strategy for Aflatoxin B₁ management. Fifty-six bacteria were isolated using a media amended with coumarin as a sole carbon source. Seven strains showed more than 70% reduction of AFB₁ in liquid culture media. Among them, isolate CFR1 reduced Aflatoxin B₁ by 94.7%, and it was selected for further studies. CFR1 was identified as Bacillus licheniformis CFR1, by biochemical characterization and 16S rRNA gene sequencing. The cell-free supernatant of B. licheniformis CFR1 was able to degrade AFB₁ efficiently than the cell lysate. The degradation of AFB₁ was examined using High-Performance Thin Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Electron spray ionization-Mass Spectrometry (ESI-MS). The optimal temperature, time, and pH of the medium for the maximum degradation of Aflatoxin B₁ were found to be 37 °C, 24 h and 7, respectively. Furthermore, Ames test for mutagenicity showed that when treated with B. licheniformis CFR1 extracellular fraction coincided with the loss of Aflatoxin B₁ mutagenicity. To the best of our knowledge, this is the first study that shows more than 90% degradation of AFB₁ by B. licheniformis. Thus, B. licheniformis CFR1 might be an excellent candidate for bioremediation and detoxification of Aflatoxin B₁ from both field and food matrices.     

Multiple mycotoxin co-occurrence in maize grown in three agro-ecological zones of Tanzania

In this study, the co-occurrence of multiple mycotoxins in maize kernels collected from 300 households' stores in three agro-ecological zones in Tanzania was evaluated by using ultra high performance liquid chromatography/time-of-flight mass spectrometry (TOFMS) with a QuEChERS-based procedure as sample treatment. This method was validated for the analysis of the main eleven mycotoxins of health concern that can occur in maize: aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), aflatoxin G₁ (AFG₁), aflatoxin G₂ (AFG₂), ochratoxin A (OTA), deoxynivalenol (DON), fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), HT-2 toxin, T-2 toxin and zearalenone (ZEN). From each zone one major maize producing district for home consumption was chosen and 20 villages for each district were randomly selected for sampling. All mycotoxins of health concern, except for T-2 toxin, were detected in the maize samples. Particularly high levels of AFB₁ (50%; 3–1,081 μg kg⁻¹), FB₁ (73%; 16–18,184 μg kg⁻¹), FB₂ (48%; 178–38,217 μg kg⁻¹) and DON (63%; 68–2,196 μg kg⁻¹) were observed. Some samples exceeded the maximum limits set in Tanzania for aflatoxins or in European regulations for other mycotoxins in unprocessed maize. Eighty seven percent of samples were contaminated with more than one mycotoxin, with 45% of samples co-contaminated by carcinogenic mycotoxins, aflatoxins and fumonisins. Significant differences in contamination pattern were observed among the three agro-ecological zones. The high incidence and at high levels (for some) of these mycotoxins in maize may have serious implications on the health of the consumers since maize constitute the staple food of most Tanzanian population. Effective strategies targeting more than one mycotoxin are encouraged to reduce contamination of maize with mycotoxins.     

Degradation and detoxification of AFB1 by Staphylocococcus warneri,Sporosarcina sp. and Lysinibacillus fusiformis

Effects associated with aflatoxins (AFs), principally aflatoxin B₁ (AFB₁) have necessitated strategies to eliminate their occurrence in commodities along the food chain. This study therefore, investigated the AFB₁ biodegradation ability of Staphylococcus warneri, Sporosarcina sp. and Lysinibacillus fusiformis liquid cultures and cell lysates (disrupted in the presence or absence of protease inhibitors to obtain lysates). These were incubated with AFB₁ (2.5 μg/mL) for 3, 6, 12, 24 and 48 h. AFB₁ degradation was subsequently monitored on high performance liquid chromatography (HPLC) and results indicated that after 48 h, % AFB₁ degradation by the liquid cultures of Lysinibacillus fusisormis, S. warneri and Sporosarcina sp. were 61.3, 47.7 and 46.9%, respectively. After 12 h of incubation, a 100% AFB₁ degradation was observed for all protease inhibited lysates tested. To establish toxicity of the AFB₁ biotransformed products, results from a cytotoxicity study against human lymphocytes demonstrated that the products exhibited significantly (p ≤ 0.05) lower cytotoxic effect compared to the parent AFB₁. From this study, it can be deduced that the mechanism of AFB₁ degradation was enzymatic and that protease inhibition of cells before disruption, could increase this enzymatic activity. Conclusively, the potential of these lysates as a biotechnological approach towards decontaminating AFB₁ is promising.     

Cross-linked chitosan polymers as generic adsorbents for simultaneous adsorption of multiple mycotoxins

The primary objective of this study was to synthesize three types of cross-linked chitosan polymers and further investigate their adsorption capability for multiple mycotoxins, including aflatoxin B₁ (AFB₁), ochratoxin A (OTA), zearalenone (ZEN), fumonisin B₁ (FB₁), deoxynivalenol (DON) and T-2 toxin (T2). Among these synthetic adsorbents, cross-linked chitosan-glutaraldehyde complex presented the highest adsorption capability for AFB₁ (73%), OTA (97%), ZEN (94%) and FB₁ (99%), but no obvious adsorption for DON and T2 (<30%). The effect of various incubation conditions (contact time, dosage and pH) was also studied. Subsequently, the experimental data were fitted to Langmuir, Freundlich and Hill models. The best fitting model to describe AFB₁ and FB₁ adsorption was Langmuir model (R² ≥ 0.99), with the theoretical maximum adsorption amounts of 5.67 mg/g for AFB₁ and 15.7 mg/g for FB₁. The Hill model was the best model for OTA and ZEN adsorption (R² > 0.98), with the predicted maximum adsorption amounts were 24.8 mg/g for OTA and 9.18 mg/g for ZEN. In addition, the adsorption capability of adsorbent for the simultaneous presence of multiple mycotoxins was also evaluated in buffer system and simulated gastrointestinal condition. The results indicated that the coexisted multiple mycotoxins didn't affected the adsorption capability of adsorbent, whereas the adsorption amounts of toxins were decreased by some gastrointestinal components. The findings of this research suggest that chitosan–glutaraldehyde complex has the potential to be applied as multitoxin adsorbent material for reducing the combined adverse effect of multiple mycotoxins on humans and animals.     

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).     

Determination of multi-mycotoxin occurrence in maize based porridges from selected regions of Tanzania by liquid chromatography tandem mass spectrometry (LC-MS/MS), a longitudinal study

Residents of certain areas of Tanzania are exposed to mycotoxins through the consumption of contaminated maize based foods. In this study, 101 maize based porridge samples were collected from villages of Nyabula, Kikelelwa and Kigwa located in different agro-ecological zones of Tanzania. The samples were collected at three time points (time point 1, during maize harvest; time point 2, 6 months after harvest; time point 3, 12 months after harvest) over a 1-year period. Ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was used to detect and quantify 9 mycotoxins: aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), aflatoxin G₁ (AFG₁), aflatoxin G₂ (AFG₂), fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), deoxynivalenol (DON), ochratoxin A (OTA) and zearaleneone (ZEN) in the samples following a QuEChERS extraction method. Eighty two percent of samples were co-contaminated with more than one group of mycotoxins. Fumonisins (FB₁ + FB₂) had the highest percentage occurrence in all 101 samples (100%) whereas OTA had the lowest (5%). For all three villages the mean concentration of FB₁ was lowest in samples taken from time point 2. Conversely, In Kigwa village there was a distinct trend that AFB₁ mean concentration was highest in samples taken from time point 2. DON concentration did not differ greatly between time points but the percentage occurrence varied between villages, most notably in Kigwa where 0% of samples tested positive. ZEN occurrence and mean concentration was highest in Kikelelwa. The results suggest that mycotoxin contamination in maize can vary based on season and agro-ecological zones. The high occurrence of multiple mycotoxins found in maize porridge, a common weaning food in Tanzania, presents a potential increase in the risk of exposure and significant health implications in children.     

Aflatoxin B1 degradation by culture and lysate of a Pontibacter specie

The presence of aflatoxin B₁ (AFB₁) along the food chain poses a significant threat, thus propelling the need for an effective approach to control it. This study was therefore, aimed at investigating AFB₁ degradation of liquid cultures and lysates of an isolated Pontibacter sp. (VGF1). Liquid cultures, lysed bacterial cells in the absence (uninhibited lysates) and presence of protease inhibitors (protease inhibited lysates) were respectively incubated with AFB₁ for 3, 6, 12, 24 and 48 h. AFB₁ degradation was monitored during this period on high performance liquid chromatography (HPLC) and results obtained revealed that after 6 h of incubation, the protease inhibited (PI) lysates yielded a 65% AFB₁ degradation, whereas after 12 h, no residual AFB₁ was detected. Conversely, after 48 h of incubation, a significantly (p≤0.05) lower AFB₁ degradation of 50 and 36% by the liquid culture and uninhibited lysate, respectively, were noted. It was further confirmed that the degradation mechanism was enzymatic. Data from cytotoxicity studies against human lymphocytes further demonstrated that extracts of biotransformed AFB₁ were less toxic when compared to that of AFB₁. Findings from this study have demonstrated an alternative approach for the decontamination and biocontrol of AFB₁ in various agricultural commodities.     

A survey of mycotoxins in random street-vended snacks from Lagos,Nigeria, using QuEChERS-HPLC-MS/MS

A survey in African snacks was carried out in order to evaluate the intake of 23 mycotoxins. The African snack samples were purchased from street vendors within Lagos metropolis (Nigeria) and evaluated for the presence of 23 mycotoxins using a modified QuEChERS procedure coupled with liquid chromatography-triple quadrupole linear ion trap mass spectrometer. The snacks included akara, baked coconut, coconut candy, donkwa, groundnut cake (kulikuli), lafun, milk curd (wara), fresh and dried tiger-nuts, and yam flour. Only three mycotoxins were detected in 23.8% of the studied snacks, and at concentrations ranging from 6 to 54 μg kg⁻¹. The concentrations of aflatoxin B₁ (AFB₁) and AFB₂ reached 23 μg kg⁻¹ and 3 μg kg⁻¹, respectively. Moreover a sample of baked coconut contained α-zearalenol (α-ZOL), which was up to 54 μg kg⁻¹ in coconut candy. As considers prevalence, aflatoxins and α-ZOL were not detected in lafun and groundnut-based snacks (donkwa and kulikuli), whereas each of the three mycotoxins contaminated 12.5% (1/8) of the coconut-based samples. This is the first report of α-ZOL in cassava and coconut, and their products. AFB₁ and total aflatoxins (TAFs) concentrations exceeded the maximum allowable limit recommended by National Agency for Food and Drug Administration and Control Nigeria (NAFDAC) in one sample of baked coconut (AFB₁ = 23 μg kg⁻¹ and TAFs = 26 μg kg⁻¹) and donkwa (AFB₁ = 19 μg kg⁻¹ and TAFs = 21 μg kg⁻¹).     

Degradation of AFB1 in aqueous medium by electron beam irradiation: Kinetics,pathway and toxicology

The degradation study of aflatoxin B₁ (AFB₁) in aqueous medium was performed under electron beam irradiation (EBI) at various AFB₁ initial concentrations. It has been proven that the degradation of AFB₁ in the selected ranges of concentrations follows pseudo first-order reaction kinetics well (R² > 0.95). Five degradation products of AFB₁ in aqueous solution were identified by UPLC-Q-TOF/MS, and the possible degradation pathway was proposed. The Ames and cytotoxicity tests were employed to evaluate the toxicity of the AFB₁ degradation products in aqueous solution, and the results indicated that the mutagenicity and cytotoxicity of EBI treated samples decreased significantly compared with that of untreated samples, but were not completely disappeared. The study provided clues involving the application of EBI methods in AFB₁ decontamination.     

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.     

Aflatoxin contamination of dried red chilies: Contrasts between the United States and Nigeria,two markets differing in regulation enforcement

Dried red chilies are among the world’s most consumed spices. From farm to fork, chilies go through cropping, harvest, drying, processing and storage. Chilies are susceptible to infection by aflatoxin producing fungi and subsequent contamination by aflatoxins at every stage. Aflatoxins are highly regulated, hepatotoxic carcinogens produced by fungi in Aspergillus section Flavi. The current study examined prevalence of aflatoxin B₁ (AFB₁) in chilies from markets across the United States (US) and Nigeria, and determined predisposition of chilies to aflatoxins post-harvest. Aflatoxin B₁ was detected in 64% chilies from US markets (n = 169), and 93% of Nigerian chilies (n = 55) with a commercial lateral flow assay (Limit of Detection = 2 μg/kg). Two percent of US samples exceeded the aflatoxin regulatory limit of 20 μg/kg, while the highest concentration detected was 94.9 μg/kg. Aspergillus spp. could be recovered only from 40% of samples from the US, and aflatoxin levels did not correlate with quantities of Aspergillus section Flavi (Colony Forming Units g⁻¹), suggesting fungi associated with chilies in US markets were killed during processing. Both average AFB₁ concentrations and fungal quantities were significantly higher (p < 0.01) in Nigerian chilies. The most contaminated sample contained 156 μg/kg AFB₁. Aflatoxin concentrations in Nigerian chilies increased as an exponential function of the quantities of Aspergillus section Flavi (r² = 0.76). Results indicate that high rates of chili consumption may be associated with unacceptable aflatoxin exposure.     

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.     

Structure elucidation and toxicity analyses of the degradation products of aflatoxin B1 by aqueous ozone

Earlier researches showed that aflatoxin B₁ (AFB₁) can be effectively degraded using ozonation in the aqueous systems. However, the degradation products have not been identified until today. In this article, the degradation products of AFB₁ were analyzed using ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometers (UPLC Q-TOF MS). Under our experimental conditions, six key degradation product structures and possible paths for generating fragment ions were proposed. The proposals were based on the low mass error, high match rate in degradation product speculation, and possible molecular formulas that could be obtained using UPLC Q-TOF MS. UPLC revealed the existence of degradation products, whereas Q-TOF MS exposed the structures of the fragment ions for AFB₁ and its degradation products. Due to the conjugate addition reaction on the double bond of the terminal furan ring for AFB₁, the toxicity of the degradation products was significantly decreased compared with that of AFB₁ through the toxicity analysis of the degradation products according to the structure–activity relationship. The results showed that aqueous ozone treatment was an effective method for degrading AFB₁.     

Aflatoxin B1 and ochratoxin A in breakfast cereals from athens market: Occurrence and risk assessment

A method for aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) determination in breakfast cereals is described using a simultaneous methanolic-aqueous extraction followed by immunoaffinity columns clean-up step and High Pressure Liquid Chromatography (HPLC) with Fluorescence Detector (FD). Recoveries were found to be 78% and 83% for AFB₁ and OTA, respectively, while the detection limit (DL) was 0.02 ng g^?1 for both mycotoxins. Both determinations were applied in fifty five samples of breakfast cereals purchased from Athens market. Results revealed the presence of AFB₁ in 56.3% of the samples examined (mean 1.42 ng AFB₁ g^?1). Seven samples (median 3.5 ng AFB₁ g^?1) were found to be contaminated at levels higher than the EU limit (2 g g^?1). OTA was detected in 60% of the samples (mean 0.18 ng g^?1). Nineteen samples were found to be contaminated by both mycotoxins. In addition in the present study the daily exposure to AFB₁ and OTA is discussed.     

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.     

Evaluation of latex agglutination inhibition reaction test for rapid detection of aflatoxin B1

Aflatoxin B₁ (AFB₁), a mycotoxin mainly produced by some Aspergillus species, has been found in a wide range of agricultural products. To avoid the risk of AFB₁ consumption, many agricultural commodities, foods and feeds should be analyzed and a rapid and non-instrumental method for the detection of AFB₁ is needed. In this study, a rapid, inexpensive and user-friendly latex agglutination inhibition reaction test (LAIRT) for on site testing of AFB₁ had been established. At first, carboxylated polystyrene latex particles (CPLP) were prepared by soap-free polymerization and sensitized with aflatoxin B₁ oxime-BSA (AFB₁O-BSA) for the detection of AFB₁. In LAIRT, the agglutination reaction with AFB₁O-BSA-sensitized CPLP and anti-AFB₁ antiserum mixture was inhibited by 5 ng/mL AFB₁ and the analysis time for 6 samples on one glass slide was less than 10 min. Subsequently, 10 rice and peanut samples were analyzed by LAIRT and ELISA, and the results showed that 1 rice sample and 2 peanut samples were positive and in agreement with those of ELISA. However, the results could be obtained more rapidly by LAIRT than ELISA. This easy and rapid LAIRT might be useful for screening AFB₁ of agricultural commodities, foods and feeds in the field.     

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.     

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.     

Development of an aptasensor for the fast detection of Versicolorin A

Aflatoxins (AFs) are secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus. The molds may contribute to pre-harvest aflatoxin contamination of susceptible crops. For the customer and food producer, a predictive model for aflatoxin detection is very desirable. Versicolorin A (VerA), which is the first precursor in the pathway of aflatoxin B₁ (AFB₁) biosynthesis, shares similar toxic group with the furofuran structure in aflatoxin B₁. VerA exhibits a much lower teratogenic toxicity than AFB₁ and may be used as a predictive indicator for aflatoxin B₁ contamination of storage crops. Therefore, the development of a fast detection method for VerA is important. One of the randomly computer-generated aptamers of VerA was confirmed by isothermal titration calorimetry with K_d = 9.26 × 10⁻⁶ mol l⁻¹. In addition, a simple and sensitive label-free aptasensor was developed for the electrochemical detection of VerA. According to the results from differential pulse voltammetry (DPV), a linear relationship existed between the log conc. of VerA (ranged from 0.01 to 100 ng ml⁻¹) and the current (△I_p) with a limit of detection (LOD) of 10 pg ml⁻¹. The resulting aptasensor exhibited good reproducibility for detecting VerA and stability after storage for 15 days at 4 °C with acceptable anti-interference against ZEN, OTA, DON, and FB₁. When used in corn samples, the recoveries of VerA were determined to be in the range of 81.3%–104.4 %. Although with some intercross, result suggests that the obtained aptamer for VerA is potentially used as a sorbent for the preparation of solid-phase-extraction procedure to clean up food samples in conjunction with high-performance liquid chromatography analysis.