Predominant mycobiota and aflatoxin content in Brazil nuts

The Brazil nut (Bertholletia excelsa) is an economically important product to the Brazilian Amazon. Currently, its marketing is being affected by the high incidence of aflatoxins (AF) produced by potentially aflatoxigenic fungi associated with its seeds. In this context, the purpose of this study was to determine which part of the nut contributes to contamination by aflatoxins and to identify the mycobiota in Brazil nut samples. Unshelled and shelled nuts were analyzed by measuring the total count of filamentous fungi (Aspergillus sections Flavi, Nigri and Circumdati) in sanitised and non-sanitised treatments. The isolates identified as Aspergillus section Flavi, the major producers of AF, were plated for determination of their aflatoxigenic potential. To perform the AF analysis, samples of Brazil nuts were treated separately. The AF from the shell and kernel were extracted by chloroform and analysed by the HPLC-FD system in isocratic mode. The Aspergillus section Flavi count was 21.67% lower. The production of AF by the isolated fungi was 30% for sanitised and 23.8% for non-sanitised samples. The concentrations obtained of AFB₁ and AFG₁ were higher than those of AFB₂ and AFG₂. The AFB₁ concentrations of shelled nuts and shell samples were 35.0 and 1.78???g/kg, respectively. AFB₂ and AFG₂ were detected only in shelled nut samples. The HPLC-FD analysis presented limits of detection (LOD) and quantification (LOQ) of 0.2 and 0.4???g/kg, respectively.     

Determination of aflatoxin risk components for in-shell Brazil nuts

A study was conducted on the risk from aflatoxins associated with the kernels and shells of Brazil nuts. Samples were collected from processing plants in Amazonia, Brazil. A total of 54 test samples (40?kg) were taken from 13 in-shell Brazil nut lots ready for market. Each in-shell sample was shelled and the kernels and shells were sorted in five fractions: good kernels, rotten kernels, good shells with kernel residue, good shells without kernel residue, and rotten shells, and analysed for aflatoxins. The kernel?:?shell ratio mass (w/w) was 50.2/49.8%. The Brazil nut shell was found to be contaminated with aflatoxin. Rotten nuts were found to be a high-risk fraction for aflatoxin in in-shell Brazil nut lots. Rotten nuts contributed only 4.2% of the sample mass (kg), but contributed 76.6% of the total aflatoxin mass (µg) in the in-shell test sample. The highest correlations were found between the aflatoxin concentration in in-shell Brazil nuts samples and the aflatoxin concentration in all defective fractions (R ^2?=?0.97). The aflatoxin mass of all defective fractions (R ^2?=?0.90) as well as that of the rotten nut (R ^2?=?0.88) were also strongly correlated with the aflatoxin concentration of the in-shell test samples. Process factors of 0.17, 0.16 and 0.24 were respectively calculated to estimate the aflatoxin concentration in the good kernels (edible) and good nuts by measuring the aflatoxin concentration in the in-shell test sample and in all kernels, respectively.     

Determination of aflatoxin risk components for in-shell Brazil nuts

A study was conducted on the risk from aflatoxins associated with the kernels and shells of Brazil nuts. Samples were collected from processing plants in Amazonia, Brazil. A total of 54 test samples (40 kg) were taken from 13 in-shell Brazil nut lots ready for market. Each in-shell sample was shelled and the kernels and shells were sorted in five fractions: good kernels, rotten kernels, good shells with kernel residue, good shells without kernel residue, and rotten shells, and analysed for aflatoxins. The kernel:shell ratio mass (w/w) was 50.2/49.8%. The Brazil nut shell was found to be contaminated with aflatoxin. Rotten nuts were found to be a high-risk fraction for aflatoxin in in-shell Brazil nut lots. Rotten nuts contributed only 4.2% of the sample mass (kg), but contributed 76.6% of the total aflatoxin mass (μg) in the in-shell test sample. The highest correlations were found between the aflatoxin concentration in in-shell Brazil nuts samples and the aflatoxin concentration in all defective fractions (R(2)=0.97). The aflatoxin mass of all defective fractions (R(2)=0.90) as well as that of the rotten nut (R(2)=0.88) were also strongly correlated with the aflatoxin concentration of the in-shell test samples. Process factors of 0.17, 0.16 and 0.24 were respectively calculated to estimate the aflatoxin concentration in the good kernels (edible) and good nuts by measuring the aflatoxin concentration in the in-shell test sample and in all kernels, respectively.     

Mycoflora and aflatoxin contamination in shelled pistachio nuts

A total of 143 pistachio nut samples collected during harvest, storage and processing were examined for mould growth and aflatoxin production. The mould count was in the range of 10³?10⁴ cfu g^?1 and 10⁵?10⁶ cfu g^?1 for the harvest and storage samples, respectively. The growth of Aspergillus flavus was 38-5-39-5% on the surface of the shells and 6–16% on the kernels without aflatoxin production. The contamination level of A flavus varied among samples collected from different regions. Peeling off the soft shell of pistachio nuts by hand reduced the contamination risk of A flavus to kernels. The predominant flora on stored pistachio nuts were Aspergillus, Penicillium, Cladosporium, Rhizopus, while the genera of Ulocladium, Trichothecium, Aureobasidium and Eurotium were less frequent. Thirty-five percent of the A flavus isolates produced aflatoxins on synthetic media.     

坚果和干果中黄曲霉毒素的污染、检测与控制

坚果和干果在生产、加工和储藏过程中易受到黄曲霉毒素污染。黄曲霉毒素是一类毒性极强的真菌毒素, 其中黄曲霉毒素B1污染最普遍、毒性最强, 其易引发急、慢性毒性, 致癌、致畸、免疫抑制等多种危害, 严重威胁果品安全, 危害人类健康。坚果和干果中黄曲霉毒素污染状况的阐释能够更加明晰其在不同果品中的污染差异, 为有针对性地控制毒素污染奠定了基础。坚果和干果中黄曲霉毒素的准确检测是分析其污染状况的前提。另外, 如何防控黄曲霉毒素在坚果和干果中的污染, 降低黄曲霉毒素对人类健康的影响, 一直是研究的重点。本文对国内外坚果和干果中黄曲霉毒素的污染情况及限量标准, 黄曲霉毒素的检测方法和防控措施进行了综述, 以期为坚果和干果中黄曲霉毒素的污染控制提供理论基础和指导。     

Consumers' ability to discriminate aflatoxin-contaminated Brazil nuts

The objectives of the study were to investigate the extent to which consumers can separate nuts with a high content of aflatoxin from sound nuts, and whether sorting results can be improved by information or whether they are affected by certain factors. A test panel consisting of 100 subjects was asked to crack 300 g Brazil nuts and to sort the nuts into those they considered edible and inedible. The test showed that consumers can, on current behaviour, discriminate aflatoxin-contaminated Brazil nuts to a significant extent. The median and the 95th percentile of the total concentrations of aflatoxins (B₁, B₂, G₁, G₂) in the samples before sorting were 1.4 and 557 µg kg^-1, respectively, and in the edible fractions after sorting 0.4 and 56 µg kg^-1, respectively. Given that levels of aflatoxins before sorting exceed either 2 µg aflatoxin B₁ kg^-1 or totally 4 µg aflatoxins kg^-1, there was no effect of aflatoxin concentrations before sorting on the probability of exceeding these thresholds in the edible fraction. This means that similar sorting results were obtained for samples with aflatoxin levels exceeding either of the two thresholds, irrespective of if the thresholds were exceeded with a few µg kg^-1 or up to more than 1000 µg kg^-1. None of the tested factors (such as sex, age, level of education, ethnic background or knowledge of mycotoxins) had any effects on the probability of exceeding either of the two aflatoxin thresholds.     

Determination of aflatoxin levels in nuts and their products consumed in South Korea

A total of 85 nuts and their products marketed in South Korea were assessed for aflatoxins using a monitoring scheme consisting of enzyme-linked immunosorbent assay (ELISA) for rapid screening, high performance liquid chromatography (HPLC) for quantification and LC–mass spectrometry (MS) for confirmation. Thirty-one out of 85 samples gave ELISA readings above 0.06 and were screened as possible positive samples. Aflatoxin contents of possible positive samples were determined using HPLC with a detection limit of 0.08–1.25 μg/kg and a quantification limit of 0.15–2.50 μg/kg. Nine samples including 1 raw peanut, 4 roasted peanuts, 2 peanut butters, 1 pistachio and 1 seasoned assorted nut were contaminated with aflatoxins (10.6% of incidence), ranging in various levels up to 28.2 μg/kg. LC–MS analysis on contaminated samples revealed that peaks eluting at 4.4, 5.2, 9.1 and 11.9 min were confirmed as aflatoxin G₁, aflatoxin B₁, aflatoxin G₂ and aflatoxin B₂, respectively.     

Brazil nuts and associated health benefits: A review

Epidemiological studies have shown an inverse relationship between nut intakes and chronic diseases such as cardiovascular diseases and cancers. The composition of lipids, minerals, and phytochemicals, and their associated health functions in Brazil nuts are critically reviewed. The nuts have high nutritive food value containing 60–70% oil and 17% protein. Brazil nuts contain abundant dietary antioxidants, especially selenium (Se). One single Brazil nut provides 160% of the US Recommended Daily Allowance (RDA) of selenium - perhaps the best source of Se from plant-based foods. Brazil nuts possess phenolics and flavonoids in both free and bound forms and are rich in tocopherol, phytosterols, and squalene. These compounds' possible beneficial effects are due to their antioxidant and antiproliferative activities, which are linked to a reduced risk for developing atherosclerosis and cancer.     

Aflatoxin production by Aspergillus flavus in Brazil nuts

Experiments were conducted to evaluate the effects of relative humidity (r.h.; 75%, 80%, 85%, 97%) and temperature (10, 13, 15, 25, 30 °C) on aflatoxin production in previously dried (3.5% moisture content; m.c.) Brazil nuts. Initially Aspergillus spp. were isolated from the surfaces of whole in-shell (WIS) Brazil nuts imported from Peru using A. flavus and A. parasiticus agar (AFPA). Isolates were subsequently screened for aflatoxin production using yeast extract sucrose medium. Total aflatoxin (B₁+B₂+G₁+G₂+M₁) was analyzed using an immunoassay technique while the presence of aflatoxin was confirmed using thin-layer chromatography. The surface of shelled half-nuts (simulating damaged or trimmed nuts), shelled whole (SW) nuts, and WIS nuts following a chlorine wash and water rinse, served as sites for inoculation (10 μl; 10⁵/ml) using an aflatoxigenic isolate. Maximum concentrations of total aflatoxin and B₁ were detected in nuts stored at 97% r.h. and at temperatures of 25–30 °C. Shelled half-nuts contained the highest total (6817 ng/g) and B₁ (4483 ng/g) aflatoxin. WIS nuts contained the least total and B₁ toxin with maximum concentrations of 93 and 49 ng/g, respectively. Aflatoxin was not detected (detection limit of 1.75 ng/g) in nuts maintained at either 10 °C (97% r.h.) or at 30 °C (75% r.h.) for up to 60 d. Maximal moisture contents (%) and water activity values (a_W) for nuts stored at these conditions were 4.50 and 0.78, and 9.14 and 0.92, respectively. Results of this study indicate that the limiting moisture content and a_W values required to control aflatoxin production () in SW and WIS stored at 30 °C for up to 60 d are 4.5, 0.68, 5.0, and 0.75, respectively. Overall, increasing the relative humidity and temperature during storage resulted in an increase in aflatoxin and these were shown to be the most significant variables influencing toxin production in Brazil nuts.     

Moisture-equilibrium relative humidity relationships in pistachio nuts with particular regard to control of aflatoxin formation

Adsorption and desorption isotherms have been determined both manometrically and by weight equilibration for Turkish pistachio nut kernel, shell and hull. Comparison of the calculated and experimentally determined adsorption isotherms for whole nuts showed good correlation. Nuts inoculated with Aspergillus flavus conidia were equilibrated to various ERH levels and stored in controlled environment cabinets at 28°C. Competitive growth of xerophilic strains of A. amstelodami prevented growth and aflatoxin production by the A. flavus at ERHs of 86% and below. At 88% ERH marked aflatoxin production occurred but competition was observed between the A. flavus and A. niger. In sealed containers metabolic moisture from growth of A. amstelodami raised the ERH from the initial 85% and permitted toxin production by A. flavus. The results are discussed in relation to post-harvest handling and storage of pistachio nuts.     

Brazil nuts: Benefits and risks associated with contamination by fungi and mycotoxins

The monotypic type genus Bertholletia produces commercially nutritionally harvested edible seeds, Brazil nuts. It is an important product from the Amazon forest in the food production chain, with a 2008 annual world production of 78,000 tonnes, being Brazil responsible for approximately 40% of it. Although there are beneficial nutritional properties, the prevailing mycobiota of Brazil nuts include fungi that are producers of aflatoxins, such as Aspergillus flavus, A. parasiticus and A. nomius. Aflatoxins have deleterious effects in consumption considering the global distribution chain, affecting major exporting countries. The present review is focused on the importance of Brazil nuts for the Amazon rainforest, emphasizing on the social and environmental impact of its production, on the mycobiota contamination of seeds, and on the presence of mycotoxins and related food safety aspects.     

Phytosterols and tocopherols content of pulps and nuts of Brazilian fruits

Fruits and nuts from the North and Northeast regions of Brazil were collected to determine their phytosterol and tocopherol content. The species studied were Cotia nut (Aptandra spruceana M.), Brazil nut (Bertholletia excelsa H.B.K.), Mucajá (Couma rigida M.), Red Açaí (Euterpe oleracea M.), Inajá (Maximiliana maripa D.), Jenipapo (Genipa Americana L.), Buriti (Mauritia flexuosa L.) and Uxi (Endopleura uchi C.). Phytosterols were analyzed by GC–FID using β-cholestanol as an internal standard, while tocopherols were determined by RP-HPLC-DAD. The pulps of Mucajá (26–236 mg 100 g^–1), Inajá (119–285 mg 100 g^–1) and Jenipapo (216 mg 100 g^–1) showed the highest total phytosterol contents. Considering α-tocopherol equivalents, the pulps of Buriti (346.72 μg g^–1) and Uxi (200.92 μg g^–1) contained the highest vitamin E activity. Therefore, the results indicate that these fruits and nuts have great potential to be cultivated and marketed as alternative dietary sources for these bioactive compounds.     

Evaluation of aflatoxin decontaminating by two strains of Saccharomyces cerevisiae and Lactobacillus rhamnosus strain GG in pistachio nuts

In this study, the surface binding ability of Saccharomayces cerevisiae and Lactobacillus rhamnosus GG (LBGG) to aflatoxin in pistachio nuts was compared. Results showed that Saccharomayces cerevisiae and Lactobacillus rhamnosus strains had aflatoxin binding ability of 40% and 35% with initial concentration of 10 ppb and 70% and 60% with initial concentration of 20 ppb aflatoxin, respectively. Acid treatment increased this ability for yeast and bacterium to 60% and 85% in first concentration and 73% and 90% for second concentration of aflatoxin, respectively. Also, heat treatment could raise surface binding of yeast to 55% and 75% for two concentrations. In addition, heat condition for Lactobacillus improved binding to 85% and 90% for two concentrations of aflatoxin. Experiments showed that microorganism’s immobilisation on contaminated pistachio had no effect on qualitative characteristics of pistachio such as colour, texture and peroxide value.     

Improvement of a sample preparation procedure for multi-elemental determination in Brazil nuts by ICP-OES

Various sample preparation procedures, such as common wet digestions and alternatives based on solubilisation in aqua regia or tetramethyl ammonium hydroxide, were compared for the determination of the total Ba, Ca, Cr, Cd, Cu, Fe, Mg, Mn, Ni, P, Pb, Se, Sr and Zn contents in Brazil nuts using inductively coupled plasma optical emission spectrometry (ICP-OES). For measurement of Se, a hydride generation technique was used. The performance of these procedures was measured in terms of precision, accuracy and limits of detection of the elements. It was found that solubilisation in aqua regia gave the best results, i.e. limits of detection from 0.60 to 41.9 ng ml^?1, precision of 1.0–3.9% and accuracy better than 5%. External calibration with simple standard solutions could be applied for the analysis. The proposed procedure is simple, reduces sample handling, and minimises the time and reagent consumption. Thus, this can be a vital alternative to traditional sample treatment approaches based on the total digestion with concentrated reagents. A phenomenon resulting from levels of Ba, Se and Sr in Brazil nuts was also discussed.     

Comparison of nixtamalization and extrusion processes for a reduction in aflatoxin content

Traditional nixtamalization and an extrusion method for making the dough ( masa ) for corn tortillas that requires using lime and hydrogen peroxide were evaluated for the detoxification of aflatoxins. The traditional nixtamalization process reduced levels of aflatoxin B 1 (AFB 1 ) by 94%, aflatoxin M 1 (AFM 1 ) by 90% and aflatoxin B 1 -8,9-dihydrodiol (AFB 1 -dihydrodiol) by 93%. The extrusion process reduced levels of AFB 1 by 46%, AFM 1 by 20% and AFB 1 -dihydrodiol by 53%. Extrusion treatments with 0, 0.3 and 0.5% lime reduced AFB 1 levels by 46, 74 and 85%, respectively. The inactivation of AFB 1 , AFM 1 and AFB 1 -dihydrodiol in the extrusion process using lime together with hydrogen peroxide showed higher elimination of AFB 1 than treatments with lime or hydrogen peroxide alone. The extrusion process with 0.3% lime and 1.5% hydrogen peroxide was the most effective process to detoxify aflatoxins in corn tortillas, but a high level of those reagents negatively affected the taste and aroma of the corn tortilla as compared with tortillas elaborated by the traditional nixtamalization process.