Mycotoxins in South African foods: a case study on aflatoxin M<Subscript>1</Subscript> in milk

The contamination of cow’s milk at the farm level with aflatoxin M₁ was investigated in South Africa. Samples of feeds, forage, maize and milk were taken at nine dairy farms, and at the same time samples of the processed milk (retail milk) were collected from the respective dairies to which the farms delivered their milk. The feeds were analysed for aflatoxin B₁ and the milk samples for aflatoxin M₁ using high performance liquid chromatography (HPLC) and fluorescence detection. All milk samples from the dairy farms were positive for aflatoxin M₁, ranging from 0.02 μg/l to 1.5 μg/l. Retail milk was also frequently contaminated with AFM₁, at levels of 0.01-3.1 μg/l. High AFB₁ levels in feed materials on the farms supplying the raw milk indicate that various sources account for this contamination frequency in milk.     

Occurrence of aflatoxin M<Subscript>1</Subscript> in commercial pasteurized milk samples in Sari,Mazandaran province,Iran

The frequency and levels of aflatoxin M₁ (AFM₁) in pasteurized milk samples in Sari, located in Mazandaran province, Iran, were determined by enzyme immunoassay. Seventy-six samples of pasteurized milk from different retail stores were randomly collected over four seasons during the year 2015. AFM₁ contamination was detected in all milk samples. The mean concentration of aflatoxin M₁ was 65.8 ng/l, with a range of 11.7–106.6 ng/l. The highest AFM₁ level was detected in milk samples collected during spring. Forty-six (60.53 %) samples had AFM₁ levels that exceeded the maximum acceptable levels (50 ng/l) recommended by the European Union (EU). Comparison of these results with previously published data for AFM₁ in milk in Iran shows that the percentage of samples exceeding the EU maximum level is consistently high over the years, indicating a general problem related to AFB₁ contamination in dairy feedingstuff.     

Aflatoxins B<Subscript>1</Subscript> and M<Subscript>1</Subscript>: risks related to milk produced in Brazil

This work shows data on the occurrence of aflatoxins in milk produced in Brazil. A review of the literature on this contamination. Several studies carried out in Brazil show that levels of aflatoxin M₁ in milk are higher than the ones established by the legislation, an evidence of the lack of control and inspection of these mycotoxins. Taking into account that milk has been widely consumed as an important source of nutrients, mainly by children, it is fundamental to carry out a thorough study of the occurrence of aflatoxins and take measures to mitigate milk contamination.     

Aflatoxin M<Subscript>1</Subscript> in human breast milk in southeastern Turkey

This study was performed to determine aflatoxin M₁ (AFM₁) in human breast milk samples collected in ?anl?urfa, located in Southeastern region of Turkey, and to investigate a possible correlation between AFM₁ occurrence (frequency and levels) and sampling seasons. Human breast milk samples collected in December 2014 and in June 2015 from a total of 74 nursing women, both outpatient and inpatient volunteers in hospitals located in ?anl?urfa, Turkey, were analyzed using competitive enzyme-linked immunosorbent assay (ELISA) for the presence of AFM₁. AFM₁ was detected in 66 (89.2%) out of 74 samples at an average concentration of 19.0 ± 13.0 ng/l (min.-max., 9.6–80 ng/l). There was a statistically significant difference between December and June concerning AFM₁ levels (p < 0.05). Further detailed studies will be needed to determine the main sources of aflatoxins in food, to establish protection strategies against maternal and infant exposure to these mycotoxins.     

Aflatoxin M<Subscript>1</Subscript> levels in different marketed milk products in Nairobi,Kenya

Milk is an important source of energy and nutrients, especially for children, and in Kenya, milk consumption is higher than other countries in the region. One major concern with milk is the risks of chemical contaminants, and reports of high levels of aflatoxin M₁ (AFM₁) in milk in Kenya has been causing public health concerns. This study collected marketed milk products every month during 1 year, just as a consumer would purchase them from retailers and traders in a low-income area, and a major supermarket in a middle/high-income area. In total, 291 sampled milk products (raw, pasteurised, UHT milk, yoghurt and lala) were collected and analysed for AFM₁ using a commercial ELISA kit. More than 50% of the samples exceeded 50 ng/kg (the level allowed in the EU), but only three samples exceeded 500 ng/kg (the level allowed in the USA). Geometric mean AFM₁ level was 61.9 ng/kg in the 135 samples from the low-income area while it was 36.1 ng/kg in the 156 from the higher income area (p?<?0.001). The levels varied significantly depending on the time of year, with lowest levels of milk in January. There were also differences between manufacturers and products, with UHT milk having lower levels. There was no difference depending on the price for all dairy products, but when only including milk, higher price was associated with lower levels of AFM₁. In conclusion, this study shows that milk purchased by a consumer is likely to contain AFM₁ above 50 ng/kg, and that further research is needed to find ways to mitigate AFM₁ contamination through working with farmers and milk processors both in the formal and informal sectors.     

Determination of serum aflatoxin B<Subscript>1</Subscript>-lysine to evaluate the efficacy of an aflatoxin-adsorbing feed additive in pigs fed an aflatoxin B<Subscript>1</Subscript>-contaminated diet

In this study, serum aflatoxin B₁ (AFB₁)-lysine was determined in order to evaluate the in vivo efficacy of a hydrated sodium calcium aluminosilicate (HSCAS) in pigs fed AFB₁. Twenty-four 49-day-old crossbred barrows were maintained in individual cages and allowed ad libitum access to feed and water. A completely randomized design was used with six animals assigned to each of four dietary treatments for 21 days as follows: (A) basal diet (BD), (B) BD supplemented with 0.5 % HSCAS, (C) BD supplemented with 1.1 mg/kg AFB₁, and (D) BD supplemented with 0.5 % HSCAS and 1.1 mg/kg AFB₁. HSCAS was able to alleviate the toxic effects of AFB₁ on pigs and reduce (P < 0.05) the levels of serum AFB₁-lysine. Cumulative reductions of adduct yield values, calculated through the equation [(pg AFB₁-lysine/mg albumin) / (μg AFB₁/kg body weight)], were 53.0, 62.8, and 72.1 after 7, 14, and 21 days of oral exposure, respectively. AFB₁-lysine has potential as an AFB₁-specific biomarker for diagnostic purposes and for evaluating the efficacy of chemoprotective interventions in pigs.     

An antibody-based microarray assay for the simultaneous detection of aflatoxin B<Subscript>1</Subscript> and fumonisin B<Subscript>1</Subscript>

Advances in microsystem technology have enabled protein and nucleic acid-based microarrays to be used in various applications, including the study of diseases, drug discovery, genetic screening, and clinical and food diagnostics. Analytical methods for the detection of mycotoxins, however, remain largely based on thin layer chromatography (TLC), high pressure liquid chromatography (HPLC), or enzyme-linked Immunosorbent assay (ELISA) . The aim of our work, therefore, was to transfer an immunological assay from microtitrr plates into microarray format, in order to develop a multiparametric, rapid, sensitive and inexpensive method for the detection of mycotoxins for use in food safety applications. Microarray technology enables the fast and parallel analysis of a multitude of biologically relevant parameters. Not only nucleic acid-based tests but also peptide, antigen, and antibody assays, using different formats of microarrays, have evolved within the last decade. Antibody-based microarrays provide a powerful tool that can be used to generate rapid and detailed expression profiles of a defined set of analytes in complex samples and are potentially useful for generating rapid immunological assays of food contaminants. In this paper, we report a feasibility study of the application of antibody microarrays for the simultaneous (or independent) detection of two common mycotoxins, Aflatoxin B₁ and Fumonisin B₁. We present the development of microarray detection of aflatoxin B1 and fumonisin B1 in standard solutions with detection limits of 3 ng/ml of AFB1 and 43 ng/ml for FB1, and have developed a competitive immunoassay in microarray format for simultaneous analyses. The quality of the microarray data is comparable to data generated by microplate-based immunoassay (ELISA), but further investigations are needed in order to characterise our method more fully. We hope that these preliminary results might suggest that further research is warranted in order to develop hapten microarrays for the immunochemical simultaneous analysis of mycotoxins, as well as for other small molecules (e.g. bacterial toxins or biological warfare agents).     

Solvent-dependent transformation of aflatoxin B<Subscript>1</Subscript> in soil

To date, all studies of aflatoxin B₁ (AFB₁) transformation in soil or in purified mineral systems have identified aflatoxins B₂ (AFB₂) and G₂ (AFG₂) as the primary transformation products. However, identification in these studies was made using thin layer chromatography which has relatively low resolution, and these studies did not identify a viable mechanism by which such transformations would occur. Further, the use of methanol as the solvent delivery vehicle in these studies may have contributed to formation of artifactual transformation products. In this study, we investigated the role of the solvent vehicle in the transformation of AFB₁ in soil. To do this, we spiked soils with AFB₁ dissolved in water (93:7, water/methanol) or methanol and used HPLC-UV and HPLC-MS to identify the transformation products. Contrasting previous published reports, we did not detect AFB₂ or AFG₂. In an aqueous-soil environment, we identified aflatoxin B_2a (AFB_2a) as the single major transformation product. We propose that AFB_2a is formed from hydrolysis of AFB₁ with the soil acting as an acid catalyst. Alternatively, when methanol was used, we identified methoxy aflatoxin species likely formed via acid-catalyzed addition of methanol to AFB₁. These results suggest that where soil moisture is adequate, AFB₁ is hydrolyzed to AFB_2a and that reactive organic solvents should be avoided when replicating natural conditions to study the fate of AFB₁ in soil.     

HPLC based method for the measurement of the reduction of aflatoxin B<Subscript>1</Subscript> by bacterial cultures isolated from different African foods

The consumption of fermented foods contaminated with aflatoxin B₁ is linked to aflatoxicosis. Aflatoxicosis is a serious problem in developing countries with environmental conditions appropriate for the biosynthesis of AFB₁ byAspergillus flavus andAspergillus parasiticus. In Africa, especially in Ghana and Nigeria, there is a very high risk of liver cancer which is caused by the consumption of AFB₁-intoxicated, traditionally fermented maize and sorghum products. It is suggested that one way to diminish this health risk might be the reduction of the AFB₁ concentration in foods by bacteria. Especially bacteria used for food fermentation processes are of great importance, with a special emphasis on lactic acid bacteria which are involved in traditionally fermented African foods based on maize and sorghum.Most publications dealing with aflatoxin degradation by microorganisms describe a phosphate buffer test system for the performance of degradation experiments. In contrast to that, a test system based on physiological active bacterial and yeast cells has been developed, to assess food fermentation organisms for their ability to reduce the AFB₁ concentration in vitro. The aflatoxin B₁ concentration in test samples was quatitatively determined by HPLC.The assessment of lactic acid bacteria originating from different German and other European culture collections only showed a very slight reduction of the AFB₁ concentration from 3% to 12%. Screening experiments in which other bacterial genera and lactic acid bacteria, isolated from different African foods have been assessed, in most cases showed the same results. However, some bacterial strains, e.g. strains of the genusBacillus derived from European culture collections and strains of the genusLactobacillus isolated from African foods, caused a release of AFB₁ which was chemically bound before to components of the test medium and which therefore could not be extracted with chloroform.A process quite similar to that may happen during food fermentations. Different experiments showed that e.g. cellulose can bind AFB₁ very effectively. Cellulose and different other food components are well known to absorb AFB₁. During fermentation the cellulose and other AFB₁-absorbing components may be degraded and the AFB₁ will be released again.The only bacterial strain known as yet which is able to reduce the AFB₁ concentration in vitro and in different food comodities isNocardia corynebacteroides (formerFlavobacterium aurantiacum). Nevertheless the mechanism of this AFB₁ reduction is actually not well understood, it still has to be investigated. In the meantime several other bacterial strains, presumably from the taxonomic group of theActinomycetes could be proved to be effective reducers of the AFB₁ concentration in our in vitro test system. Because as yet no food relevant microorganism could be found, which is able to degrade AFB₁, these new strains in general offer the possibility for a genetic modification of food relevant microorganisms. This seems to be the way to come to starter cultures which are able to degrade AFB₁ during food fermentations.     

Exposure assessment and risk characterization of aflatoxin B<Subscript>1</Subscript> in Malaysia

Aflatoxin B₁ (AFB₁) was detected in 57% of the nuts and nut products marketed in Penang, Malaysia using the liquid chromatography tandem mass spectrometry. The contamination levels ranged from 0.40 to 222 μg/kg and 17 out of 128 samples (13.3%) contained AFB₁ above the European Commission permitted level (2 μg/kg). Estimated dietary exposure of AFB₁ in nuts and nut products were 0.36 ng per kg body weight and day and 8.89 ng per kg body weight and day, representing the low and high-level of exposure, respectively. Dose-response modelling resulted in benchmark dose lower confidence limit (BMDL₁₀) values of 0.305 ng per kg body weight and day, with the best fitted from the log-logistic model. The derived margin of exposure (MoE) values ranged from 34 to 847 suggested that AFB₁ would be of public health concern and might reasonably be considered as a high priority for risk management actions.     

An enzymatic method for the determination of hemoglobinA<Subscript>1C</Subscript>

Fructosyl peptide oxidase is a flavoenzyme that catalyzes the oxidative deglycation of N-(1-deoxyfructosyl)-Val-His, a model compound of hemoglobin (Hb)A_1C. To develop an enzymatic method for the measurement of HbA_1C, we screened for a proper protease using N-(1-deoxyfructosyl)-hexapeptide as a substrate. Several proteases, including Neutral protease from Bacillus polymyxa, were found to release N-(1-deoxyfructosyl)-Val-His efficiently, however no protease was found to release N-(1-deoxyfructosyl)-Val. Neutral protease also digested HbA_1C to release N-(1-deoxyfructosyl)-Val-His, and then the fructosyl peptide was detected using fructosyl peptide oxidase. The linear relationship was observed between the concentration of HbA_1C and the absorbancy of fructosyl peptide oxidase reaction, hence this new method is a practical means for measuring HbA_1C.     

Resolution-optimized NMR measurement of <Superscript>1</Superscript>D<Subscript>CH</Subscript>, <Superscript>1</Superscript>D<Subscript>CC</Subscript> and <Superscript>2</Superscript>D<Subscript>CH</Subscript> residual dipolar couplings in nucleic acid bases

New methods are described for accurate measurement of multiple residual dipolar couplings in nucleic acid bases. The methods use TROSY-type pulse sequences for optimizing resolution and sensitivity, and rely on the E.COSY principle to measure the relatively small two-bond ²D_CH couplings at high precision. Measurements are demonstrated for a 24-nt stem-loop RNA sequence, uniformly enriched in ¹³C, and aligned in Pf1. The recently described pseudo-3D method is used to provide homonuclear ¹H-¹H decoupling, which minimizes cross-correlation effects and optimizes resolution. Up to seven ¹H-¹³C and ¹³C-¹³C couplings are measured for pyrimidines (U and C), including ¹D_C5H5, ¹D_C6H6, ²D_C5H6, ²D_C6H5, ¹D_C5C4, ¹D_C5C6, and ²D_C4H5. For adenine, four base couplings (¹D_C2H2, ¹D_C8H8, ¹D_C4C5, and ¹D_C5C6) are readily measured whereas for guanine only three couplings are accessible at high relative accuracy (¹D_C8H8, ¹D_C4C5, and ¹D_C5C6). Only three dipolar couplings are linearly independent in planar structures such as nucleic acid bases, permitting cross validation of the data and evaluation of their accuracies. For the vast majority of dipolar couplings, the error is found to be less than ±3% of their possible range, indicating that the measurement accuracy is not limiting when using these couplings as restraints in structure calculations. Reported isotropic values of the one- and two-bond J couplings cluster very tightly for each type of nucleotide.     

Phospholipid metabolism is required for M<Subscript>1</Subscript> muscarinic inhibition of N-type calcium current in sympathetic neurons

The signal transduction cascade mediating muscarinic receptor modulation of N-type Ca²⁺ channel activity by the slow pathway has remained incompletely characterized despite focused investigation. Recently we confirmed a role for the G-protein G_q and identified phospholipase C (PLC), phospholipase A₂ (PLA₂), and arachidonic acid (AA) as additional molecules involved in N-current inhibition in superior cervical ganglion (SCG) neurons by the slow pathway. We have further characterized this signal transduction cascade by testing whether additional molecules downstream of phosphatidylinositol-4,5-bisphosphate (PIP₂) are required. The L-channel antagonist nimodipine was bath-applied to block L-current. Pretreating cells with pertussis toxin (PTX) minimized M₂/M₄ muscarinic receptor inhibition of N-current by the membrane-delimited pathway. Consistent with our previous studies, pharmacologically antagonizing M₁ muscarinic receptors (M₁Rs), G_q, PLC, PLA₂, and AA minimized N-current inhibition by the muscarinic agonist oxotremorine-M (Oxo-M). When cells were left untreated with PTX, leaving the membrane-delimited pathway intact and the same antagonists retested, Oxo-M decreased whole cell currents. Moreover, inhibited currents displayed slowed activation kinetics, indicating intact N-current inhibition by the membrane-delimited pathway. These findings indicate that the antagonists used to block the slow pathway acted selectively. PLA₂ cleaves AA from phospholipids, generating additional metabolites. We tested whether the metabolite lysophosphatidic acid (LPA) mimicked the inhibitory actions of Oxo-M. In contrast to AA, applying LPA did not inhibit whole cell currents. Taken together, these findings suggest that the slow pathway requires M₁Rs, G_q, PLC, PIP₂, PLA₂, and AA for N-current inhibition.Abbreviations AA arachidonic acid - BAPTA 1,2-bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid - BSA bovine serum albumin - DAG diacylglycerol - DEDA 7,7-dimethyleicosadienoic acid - ETYA 5,8,11,14-eicosatetraynoic acid - FPL FPL 64176 - IP₃ inositol-1,4,5-trisphosphate - L-channel L-type calcium channel - L-current L-type calcium current - LPA lysophosphatidic acid - M₁R M₁ muscarinic receptor - N-channel N-type calcium channel - N-current N-type calcium current - NMN nimodipine - OAG 1-(cis-9-octadecenoyl)-2-acetyl-sn-glycerol - OPC oleoyloxyethyl phosphorylcholine - Oxo-M oxotremorine methiodide - PIP₂ phosphatidylinositol-4,5-bisphosphate - PLC phospholipase C - PLA₂ phospholipase A₂ - PTX pertussis toxin - SCG superior cervical ganglion     

Biomolecule-functionalized magnetic nanoparticles for flow-through quartz crystal microbalance immunoassay of aflatoxin B<Subscript>1</Subscript>

A flow-through quartz crystal microbalance (QCM) immunoassay method has been developed based on aflatoxin B₁ antibody (anti-AFB₁)-functionalized magnetic core-shell Fe₃O₄/SiO₂ composite nanoparticles (bionanoparticles) in this study. To construct such an assay protocol, anti-AFB₁, as a model protein, was initially covalently immobilized onto the Fe₃O₄/SiO₂ surface, and then the functionalized nanoparticles were attached to the surface of the QCM probe with an external magnet. The binding of target molecules onto the immobilized antibodies decreased the sensor’s resonant frequency, and the frequency shift was proportional to the AFB₁ concentration in the range of 0.3–7.0 ng/ml. The regeneration of the developed immunosensor was carried out via attaching or detaching the external magnet from the detection cell. In addition, the selectivity, reproducibility, and stability of the proposed immunoassay system were acceptable. Compared with the conventional ELISAs, the proposed immunoassay system was simple and rapid without multiple labeling and separation steps. Importantly, the proposed immunoassay method could be further developed for the immobilization of other antigens or biocompounds.     

Electronic structure and bonding of the dinuclear metal M<Subscript>2</Subscript>(CO)<Subscript>10</Subscript> decacarbonyls: applications of natural orbitals for chemical valence

The nature of the chemical metal–metal bond in M₂(CO)₁₀ (M?=?Mn, Re, Tc) dinuclear decacarbonyls complexes was investigated for the first time using the natural orbital chemical valence (NOCV) approach combined with the extended transition state (ETS) for energy decomposition analysis (EDA). The optimized geometries carried out at different levels of theory BP86, BLYP, BLYPD and BP86D, showed that the latter method, i.e., BP86D, led to the best agreement with X-ray experimental measurements. The BP86D/TZP results revealed that the computed covalent contribution to the metal–metal bond are 60.5%, 54.1% and 52.0% for Mn–Mn, Re–Re and Tc–Tc, respectively. The computed total interaction energies resulting from attractive terms (ΔE _orb and ΔE _eles), correspond well to experimental predictions, based on bond lengths and energy interaction analysis for the studied complexes.     

The effect of <Emphasis Type="Italic">Baccharis glutinosa</Emphasis> extract on the growth of mycotoxigenic fungi and fumonisin B<Subscript>1</Subscript> and aflatoxin B<Subscript>1</Subscript> production

The aim of this study was to evaluate the effect of Baccharis glutinosa isolated extract on the growth of Aspergillus flavus and Aspergillus parasiticus, and their aflatoxin B₁ production; and growth of Fusarium verticillioides, and their fumonisin B₁ production. The three fungi were exposed to an antifungal fraction, designated as fraction F6-1, isolated from B. glutinosa by methanolic extraction followed by silica gel chromatography. The growth of the fungi was evaluated in kinetics of radial extension growth, kinetics of spores germination, length and diameter of hyphae, spores diameter, as well as in aflatoxin B₁ and fumonisin B₁ production. Fraction F6-1 caused radial growth inhibition of the three fungi mainly F. verticillioides. Spores germination of A. flavus and A. parasiticus was delayed in the early stage of the incubation time, although they completely germinated at 27 h. In contrast, spore germination of F. verticillioides was inhibited 87.7% up to 96 h. The lengths and diameters of hyphae, and spore diameters of the three fungi, were significantly smaller in comparison with those of the controls, and several morphological alterations were observed. Concerning aflatoxin B₁ and fumonisin B₁, fraction F6-1 did not show any inhibition effect at the concentration used. Fraction F6-1 was able to significantly inhibit the development of the three fungi, mainly F. verticillioides. The strong inhibitory effect of F6-1 on hyphae and spores suggests that it interacted with the fungi cell walls, which caused severe deformities. Nevertheless, this fraction was unable in inhibiting mycotoxin production from the three fungi at the concentration tested.     

Differences between cellular mechanisms of ATP-and noradrenaline-induced inhibition of visceral smooth muscles under conditions of selective and combined activation of M<Subscript>2</Subscript> or M<Subscript>3</Subscript> cholinoreceptors

Our studies of the role of phospholipase C in inhibitory synaptic action upon visceral smooth muscles demonstrated that, under conditions of carbachol (CCh)-induced pre-activation of cholinoreceptors, ATP-or noradrenaline (NA)-evoked relaxation of these muscles is mediated by the phospholipase C-independent pathway, while the phospholipase C-dependent pathway does not manifest itself as a mechanism that determines the inhibitory effect of the above transmitters. Under conditions of pre-activation of muscarinic cholinoreceptors, ATP-and NA-induced relaxation is continued due to activation of inositol trisphosphate (InsP₃)-sensitive receptors despite the fact that the pathway of inhibition is phospholipase C-independent. This is confirmed by complete depression of the inhibitory effects of ATP and NA against the background of CCh-induced contraction after pre-incubation of the studied preparations together with 100 μM 2-APB, a blocker of InsP₃ receptors. Selective blockings of either M₂ or M₃ cholinoreceptors are accompanied by a complete loss of the ability of the above blocker of InsP₃ receptors (2-APB) to suppress ATP-and NA-induced contraction of smooth muscles in the state of CCh-induced contraction. It can be hypothesized that, under conditions of selective pre-activation of M₂ or M₃ cholinoreceptors, the mechanisms of intracellular signalling mediating the inhibition events are modified. The InsP₃-dependent pathway that determines both adrenergic and purinergic inhibition of smooth muscles is switched off, and the inhibitory action of neurotransmitters is realized under such conditions through the InsP₃-independent pathway. Therefore, in our study we first found differences between cellular mechanisms underlying ATP-and NA-induced inhibition of smooth muscles under conditions of selective activation of either M₂ or M₃ cholinoreceptors and the mechanisms underlying the relaxing action of inhibitory neurotransmitters under conditions of combined synergistic activation of cholinoreceptors of both the above-mentioned subtypes. Neirofiziologiya/Neurophysiology, Vol. 39, No. 1, pp. 22–31, January–February, 2007.     

Exposure to aflatoxin B<Subscript>1</Subscript> in Thailand by consumption of brown and color rice

This study assessed the aflatoxin B₁ (AFB₁) intake of the Thai population through consumption of contaminated brown and color rice. A total of 240 rice samples from two harvesting periods were collected in June/July 2012 (period I) and in December 2012/January 2013 (period II) and analyzed for AFB₁ by HPLC with fluorescence detection (limit of detection (LOD)?=?0.093 ng/g). Exposure assessment was based on AFB₁ levels in rice and food intake data for rice according to Thai National Consumption. Frequency and levels of AFB₁ were higher in period I (59 %, <LOD?=?26.61 μg kg^?1) than in period II (10 %, <LOD?=?3.51 μg kg^?1). Only one sample exceeded the Thai standard limit for total aflatoxin of 20 μg kg^?1, but 12 out of 240 rice samples exceeded the European Union maximum level for AFB₁ of 2 μg kg^?1. The data showed that the quality and safety of Thai rice largely comply with the requirement for both exports and domestic consumption. According to the Thai National Consumption data, the estimated AFB₁ intake via rice consumption in period I and period II was 0.80 and 0.12 μg kg^?1 bw day^?1, respectively. The potential risk for cancer, based on the recommendation of the JECFA, was estimated to be 0.011 person/year/100,000 people at a mean consumption. Although the risk via consumption of Thai rice seems to be low, the maximum levels of AFB₁ in this staple food suggest that careful monitoring and surveillance of AFB₁ contamination in rice is essential to ensure the safety of rice.     

F<Subscript>1</Subscript>-ATPase as an auto-oscillatory system

In a relatively simple mathematical model we analyze the possibility of auto-oscillatory modes of F₁-ATPase during ATP hydrolysis.