Aflatoxins In Foods Research Articles

Legal Regulation and Analytical Method for Mycotoxin in Japan

Mycotoxins, toxic secondary metabolites produced by fungi, are present in food and feed worldwide. Acute and chronic dietary exposures can induce adverse health effects in humans and animals. Among the various mycotoxins, aflatoxins pose significant health concerns to the general public. In the early 1960s, a total of more than 100000 turkey poults died from an unknown turkey "X" disease in England. The disease was associated with Brazilian groundnut meal contaminated by Aspergillus flavus, from which aflatoxins were first isolated from the fungal culture broth. Subsequent studies revealed that aflatoxin B1 (AFB1) is the most potent carcinogen among all aflatoxins, affecting both humans and various animal species. The International Agency for Research on Cancer has classified AFB1 as a Group 1 human carcinogen. Aflatoxins are present in a wide variety of food items, including cereals, nuts, fruits, and spices. A survey conducted in Japan between 2004 and 2006 revealed that peanut products, cacao products, peppers, and Job's tears were contaminated with aflatoxins. To reduce exposure, Japan has set a regulatory limit of 10 µg/kg for total aflatoxins [sum of AFB1, aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), and aflatoxin G2 (AFG2)] for all food items. The National Institute of Health Sciences has developed official analytical methods for determining aflatoxins in foods which are used for quarantine inspection of imported foods. In this symposium, the regulations and analytical methods for aflatoxins are introduced.

Gene recognition and role of foodomics in mycotoxin control: A review

Since recognition of toxic and carcinogenic aflatoxins in Brazilian groundnut meal in 1960, much research has been done to prevent and detoxify aflatoxins in foods and feeds, identifying a variety of methods. The research has expanded to other mycotoxins. The biotic and abiotic factors favoring mycotoxin contaminations have been understood through experiments under laboratory conditions and analysis of field data. However, many gaps remain in the knowledge on mycotoxin control at the molecular level that may be useful in addressing mycotoxigenic hazards. Recognition of responsible genes in hosts and fungi and omics methods applying genomics, transcriptomics, proteomics, and metabolomics to understand mycotoxin biosynthesis at the molecular level may open new avenues to interact with plant-fungi-bacteria cross-talks, apply regulatory mechanisms in biosynthesis, and explore checks and controls addressing abiotic and biotic factors favoring mycotoxin biosynthesis. The new knowledge is expected to generate probable molecular biological mechanisms to eliminate mycotoxin biosynthesis on foods. The current level of omics knowledge requires application of research to achieve deeper understanding, aiming at new methods for mycotoxin controls and applying next-generation technologies. This review examines the current knowledge on the biosynthesis of aflatoxins, fusarium toxins, and patulin in foods and host-fungi interactions at a molecular level.

Simultaneous efficient capture of alkaloids and aflatoxins in foods using porous TiO2-assisted miniaturized solid-phase extraction via ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry

Simultaneous efficient capture of alkaloids and aflatoxins in foods using porous TiO2-assisted miniaturized solid-phase extraction via ultrahigh performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry

Physicochemical characterization and cytotoxicity assessment of sodium dodecyl sulfate (SDS) modified chitosan (SDSCS) before and after removal of aflatoxins (AFs) as a potential mycotoxin Binder

Aflatoxins in food and feed with prominent toxic effects have jeopardized public health for decades. This investigation intends to explore synthesized SDS-modified chitosan as new generation of binder for removal of aflatoxin using a straightforward ionic cross-linking approach. The primary objective of this technique was to enhance affinity and adsorption capability of SDSCS towards aflatoxins. In this context, physicochemical properties of SDSCS characterized with advanced analytical techniques such as scanning electron microscope (SEM) coupled with energy dispersive X-ray spectroscopy (EDS) and Fourier transform infrared spectroscopy (FT-IR) before and after removal of aflatoxin. In this study, effect of the pH on the adsorption of aflatoxins (6ppb) indicated that the increase in SDSCS concentration from low (0.5) to high (2 %) resulted in an increase of about 80 %, 78 % and 81 % in the adsorption percentage of AFB1, AFG1, and AFB2 & AFG2, respectively. FT-IR analysis showed the intramolecular interactions of the amine groups of chitosan and sulfate group of SDS formed a stable complex in the removal of aflatoxin that verified with appearance of three new additional peaks at 1323.50, 984.34 and 603.42 cm−1. Notably, SEM images revealed that the porous SDSCS network was filled with aflatoxin molecules supported with EDS findings. Also, in vitro cytotoxicity assessments demonstrated that SDSCS protected HepG2 cells against cytotoxic effect caused by aflatoxin (5 µM) in a concentration-dependent manner compared to the control (p<0.01). Collectively, the adsorption mechanism may involve attraction of anionic aflatoxin molecule into the interconnected pores of SDSCS complex with numerous cationic active site via hydrogen bond and van der waals force.

Determination of Total Aflatoxins in Polished Rice by Liquid Chromatography-Fluorescence Detection with Multifunctional Column Cleanup and Precolumn Derivatization: Single-Laboratory and Inter-Laboratory Validation Studies.

Aflatoxins (AFs) are toxic metabolites produced by Aspergillus spp. Because AFs are potent carcinogens in humans and animals, many countries have set regulatory limits for AFs in foods to prevent dietary exposure. From a global food safety perspective, in 2023, the Codex Alimentarius Commission established the maximum level (ML) of total AFs in certain cereals and cereal-based products, including polished rice. Therefore, validated analytical methods for AFs detection are necessary. In this study, an HPLC-fluorescence method coupled with multifunctional column cleanup and trifluoroacetic acid derivatization was developed for the determination of AFlevels in polished rice. Our method was validated in a single-laboratory study using AF-spiked materials, followed by an inter-laboratory validation study. Twelve laboratories participated in the inter-laboratory validation study, and five polished rice test samples artificially contaminated with AFswere analyzed. In a single-laboratory study, the ranges of mean recoveries of AFB1, B2, G1, G2, and total AFswere 101, 100-103, 93-96, 95-98, and 97-99%, respectively. The RSDs for within-day and between-day variations were all ≤4.4%. In the inter-laboratory validation study, the RSDs for repeatability and reproducibility were from 0.7 to 2.7% and 3.3 to 8.9% for all analytes, respectively. In response to the Codex ML and method performance criteria for AFsin polished rice, an analytical method based on HPLC-fluorescence detection was developed. All method performance parameters estimated from the test results of the single-laboratory and inter-laboratory validation studies met the criteria required by the Codex. Single- and inter-laboratory studies for the validation of an analytical method for AFlevel determination in polished rice were successfully performed. This analytical method will be suitable to determine AFlevels around the Codex ML set for polished rice.

Technologies to decontaminate aflatoxins in foods: a review

SummaryAflatoxins are toxic secondary metabolites produced by Aspergillus spp., found in staple food commodities. Aflatoxins are carcinogenic, teratogenic, and mutagenic and pose a serious threat to the health of humans. The identification and quantification of aflatoxins in foods is a major challenge to guarantee food safety. Therefore, developing feasible, sensitive, and robust methods for decontamination is paramount, with short processing time and negligible impact on quality. This review evaluates recent novel technologies for aflatoxins decontamination by physical methods (microwave heating, Gama and electron beam irradiation, pulse light and ultraviolet), chemical methods (ozone, natural plant extracts, and organic acids), and biological methods (atoxigenic Aspergillus strains, Trichoderma spp., and bacteria and yeast). The study highlights on the cutting‐edge technologies of smart packaging and artificial intelligence (AI). To achieve more efficiency and adaptability to different food matrices in aflatoxins decontamination, the study suggests integrating multiple strategies. The study also recommends integrating Partnership for Delivery (P4D) to share the responsibility to increase the chance for success and control aflatoxins in foods.

Current Strategies in Controlling Aspergillus flavus and Aflatoxins in Grains during Storage: A Review

Aspergillus flavus is a ubiquitous pathogen that can infect many foods and grains, and it produces large amounts of aflatoxins during their storage. Aflatoxins are considered highly toxic and carcinogenic to humans, and they cause great damage to crop production, food security, and human health. Thus, controlling A. flavus and aflatoxins in grains presents a great challenge to humans worldwide. Over the past decade, many strategies have been demonstrated to be useful in controlling A. flavus and aflatoxins during food storage. These methods involve physical agents, chemical agents, biological agents, etc. Some of these methods are currently used in actual production. In this review, we summarize the reported methods for controlling A. flavus and aflatoxins during food storage in the past ten years and elucidate their advantages and disadvantages. The methods discussed include irradiation technology; low oxygen atmospheres; chemical fungicides (benzalkonium chloride, iodine, ammonium bicarbonate, and phenolic and azole compounds); biological agents from plants, animals, and micro-organisms; and aflatoxin elimination methods. We expect that this review will promote the applications of current strategies and be useful for the development of novel technologies to prevent or eliminate A. flavus and aflatoxins in food and feed during storage.

Review: Sample Pretreatment and Detection Strategies for Aflatoxins in Foods

Review: Sample Pretreatment and Detection Strategies for Aflatoxins in Foods

Determination of Aflatoxins in Pig Feed. Using the Thin Layer Chromatography Technique

Aflatoxins are the most important mycotoxins in terms of food safety, due to their frequency and toxicity, they are characterized by being secondary metabolites synthesized by fungi. In order to determine the presence of aflatoxins in pork feed, five samples from different commercial companies were analyzed. The methodology used prior to the sample extraction process was the thin layer chromatography technique, observed using long-wave ultraviolet light. It could be seen in samples 1,3,4,5, the presence of Aflatoxins, toxicogenic strains of Aspergillus parasiticus, which produce aflatoxins, G1 and G2, is characterized by the fluorescence coloration, in sample 2 the presence was observed of aflatoxins toxicogenic strains of Aspergillus flavus. Toxigenic strains of Aspergillus flavus generally produce only aflatoxins B1 and B2, while toxicogenic strains of Aspergillus parasiticus produce aflatoxins B1, B2, G1 and G2. AFB1 aflatoxins have been considered obvious carcinogens in experimental animals. It is important to carry out systematic research with low-cost analytical techniques to understand the mechanisms of production and action of aflatoxins in foods, in order to prevent, monitor and control these types of toxins.

Aflatoxins in food: Prevalence, health effects, and emerging trends in its mitigation—An updated review

AbstractMycotoxins are secondary metabolites produced by a variety of fungal and mold species. One of the mycotoxin families that is often regarded as the most concerned one in human and animal feeds, is aflatoxin. Aflatoxins produced by Aspergillus species are very toxic and cause significant contamination of food supplies, resulting in serious health repercussions including carcinogenicity. To date, several regulations on aflatoxin permissible level limits have been established in a number of nations. Based on government guidelines and restrictions, consumers and food producers anticipate that aflatoxin contamination in food should not negatively impact human and animal health. Aflatoxin contamination of food commodities poses a consequential risk to humans and is also associated with substantial financial losses. Conventional mycotoxin reduction strategies involve both preventative and decontamination approaches. The present review gives an update on recent research on conventional approaches, including pre‐ and post‐harvest procedures that could potentially be used to decontaminate aflatoxin. Additionally, the current study delves into the source of contamination, global occurrence, impact on human health, and management strategies to assure food security and safety.

Metabolites and degradation pathways of microbial detoxification of aflatoxins: a review.

The degradation of aflatoxins using nonpathogenic microbes and their enzymes is emerging as a safe and economical alternative to chemical and physical methods for the detoxification of aflatoxins in food and feeds. Many bacteria and fungi have been identified as aflatoxin degraders. This review is focused on the chemical identification of microbial degradation products and their degradation pathways. The microbial degradations of aflatoxins are initiated by oxidation, hydroxylation, reduction, or elimination reactions mostly catalyzed by various enzymes belonging to the classes of laccase, reductases, and peroxidases. The resulting products with lesser chemical stability further undergo various reactions to form low molecular weight products. Studies on the chemical and biological nature of degraded products of aflatoxins are necessary to ensure the safety of the decontamination process. This review indicated the need for an integrated approach including decontamination studies using culture media and food matrices, proper identification and toxicity profiling of degraded products of aflatoxins, and interactions of microbes and the degradation products with food matrices for developing practical and effective microbial detoxification process.

Synthesis of a core-shell magnetic covalent organic framework for the enrichment and detection of aflatoxin in food using HPLC-MS/MS.

A porous magnetic covalent organic framework, Fe3O4@TPBD-TPA (terephthalaldehyde (TPA) , N, N, N', N'-tetrakis(p-aminophenyl)-p-phenylenediamine (TPBD)), was synthesized using the Schiff base reaction under mild reactionconditions. This adsorbent exhibited excellent adsorption performance for aflatoxins. The adsorption capacity of Fe3O4@TPBD-TPA for aflatoxins ranged from 64.4 to 84.4 mg/g. A magnetic solid-phase extraction combined with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method based on Fe3O4@TPBD-TPA was developed for the efficient determination of four types of aflatoxins in food samples (maize, maize oil, peanut, and peanut oil). The determination coefficients (R2) were ≥0.9972. The method exhibited detection limits ranging from 0.01 to 0.06 μg/kg and spiked recoveries of 80.0 to 113.1%. The intra-day and inter-day precision were less than 6.77%, indicating good repeatability. The adsorbent showed promising prospects for the efficient enrichment of trace amounts of aflatoxins in complex food matrices.

Aflatoxins posing threat to food safety and security in Pakistan: Call for a one health approach.

Aflatoxins are among the most important mycotoxins due to their widespread occurrence and adverse impacts on humans and animals. These toxins and/or their metabolites cannot be destroyed with cooking or boiling methods. Therefore, consumption of aflatoxin-contaminated food may lead to impaired growth, compromised immunity, stomach and liver cancer, and acute toxicity. These adverse effects along with food wastage might have detrimental consequences on a country's economy. Several studies from Pakistan reported a high prevalence of aflatoxins in food and feed commodities (Range; milk=0.6-99.4%, cereals, and grains=0.38-41%, animal feed=31-100%). Notably, Pakistan reported very high figures of impaired child growth-stunted 40.2%, wasted 17.7% and underweight 28.9%-that could be associated with the higher aflatoxin prevalence in food items. Importantly, high aflatoxins prevalence, i.e. 100%, 69% and 60.5%, in children has been reported in Pakistan. Food and feed are more prone to aflatoxin contamination due to Pakistan's hot and humid climate; however, limited awareness, inadequate policy framework, and weak implementation mechanisms are the major obstacles to effective control. This review will discuss aflatoxins prevalence, associated risk factors, adverse health effects, required regulatory regime, and effective control strategies adopting the One Health approach to ensure food safety and security.

Ensuring food safety with molecularly imprinted polymers: innovative methods for the detection of aflatoxins in food and feed samples

Aflatoxins, a group of mycotoxins, represent a heterogeneous class of secondary metabolites that pose a significant risk to food safety and public health due to their potent toxicity. Aflatoxins are widely distributed in the environment, with high levels frequently observed in hot and humid conditions. There is an ongoing development of various methods for detecting aflatoxins in food and feed samples. Herein, a review of these methods is presented with special emphasis on molecularly imprinted polymers (MIPs) as selective materials for aflatoxins’ detection. The key findings of various methods for real-time analysis of food and feed samples are presented and analyzed, providing a comparative assessment of their performance. Furthermore, the challenges and limitations of these methods are discussed, considering their commercialization prospects and real-world requirements.

Effectiveness of some aflatoxin interventions in reducing the aflatoxin contamination of maize in Ghana

The presence of aflatoxin in food and feed represents a significant health hazard to both human and animal populations, hence exerting adverse effects on the environment and the global economy. The effects of maize variety, agroecological zones, and aflatoxin control measures on aflatoxin contamination were tested using a 2x2x3 factorial in a randomised complete block design (RCBD). The treatments consisted of two agroecological zones, two varieties of maize, and three aflatoxin control measures. The aflasafe was applied at a rate of 10 kg/ha and the nicoking at 200 ml per 15 liters of water. There was no contamination of maize samples grown in the Forest zone with aflatoxins during all the treatments. There was no contamination of maize samples with aflatoxins BI, B2, G1, and G2 for all the treatments in the Savannah zone except for the hoeing method. The highest contamination of Aflatoxin G2 (AFG2) (0.19 µg/kg) was recorded by the Wangdataa maize variety using hoeing as the means of the aflatoxin control method. The highest Aflatoxin G1 (AFG1) contamination (0.45 µg/kg) was produced by maize grown in the Savannah ecozone using the hoeing method of control. In conclusion, the study showed that all the control methods gave maximum protection to the maize while growing on the field except the hoeing method in the Savannah ecological zone. The study recommends that in the Savannah zone if a farmer chooses hoeing as a method to control aflatoxins, it is advisable to complement this approach with the use of biological or chemical methods for effective aflatoxin control.

Polydiacetylene based colorimetric nano-sensor for on-spot detection of aflatoxin in food matrices and animal feed

Global food security is constantly being challenged by the prevalence of aflatoxin (AF), a carcinogenic mycotoxin found in various staple foods consumed across the globe. This challenge is further compounded by unavailability of convenient, on-field aflatoxin sensing devices. In this study, liposome sensors conjugated with polydiacetylene (PDA) have been developed for robust, low-cost, on-site quantification of aflatoxin from feed samples assisted by a convenient mobile phone-based application. Soyabean-sourced phosphatidylcholine (content ≥ 94.0%) demonstrated to be a stronger structural backbone for developed lipoid sensor and is comparatively cheaper and sustainable alternative to conventional backbones like synthetic 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC) for the preparation of based liposomes. Anti-aflatoxin antibodies conjugated with chemically activated liposomes provide a consistent chromatic response with aflatoxin. These sensors were used to detect the presence of aflatoxin in food and animal feed matrices. The detection time of the developed sensor was 5 min and can provide rugged and reliable detection of AFB1 up to the limit of 5.7 parts per billion (ppb).

Correlation between Aspergillus flavus fungal biomass and aflatoxin contamination in harvested maize: insights from Kenya and Tanzania

Controlling the occurrence of aflatoxins in foods must be accompanied by managing the fungi responsible for their production. The abundance and diversity of aflatoxin-producing Aspergillus flavus are responsible for the accumulation of these toxins in crops, posing a persistent threat to public health and the economy in tropical developing countries. A study was conducted to investigate the occurrence and level of A. flavus and relate them to aflatoxin levels in maize in Kenya and Tanzania. A total of 786 maize samples were collected during harvesting in selected areas of the two countries for analysis. The fungal abundance in the samples was measured as the amount of fungal DNA relative to maize DNA. This was accomplished by quantifying the fungal DNA using qPCR, targeting the internal transcribed spacer (ITS) gene, while the maize DNA was quantified through the alpha-tubulin gene, the two genes known to be conserved. Aflatoxins were quantified using ultra-high performance liquid chromatography, coupled with ultra-high sensitivity, ultra-fast triple quadrupole tandem-mass spectrophotometer. A. flavus was detected in 88.5% of the 786 tested samples, and the average fungal load for these samples (expressed as the log host/pathogen ratio) was 5.53. Aflatoxin occurrence was positive in 31.9% of the samples, with an average level of 2.3 ± 0.643 ppb. The study established a positive relationship between the occurrence and level of aflatoxin B1 and the presence and biomass of A. flavus, which was statistically proven. These findings emphasize the need to place substantial attention on preharvest control of A. flavus in cereal fields as an effort to control the accumulation of aflatoxin B1 in foods.

Streptomyces tetracycline’s computational behavior against polyketide synthase of aflatoxigenic fungi

produces a variety of bioactive compounds that prevent fungal growth, including aflatoxins. Aflatoxigenic fungi ( and ) are being researched concerning spp. and can prevent the spread of aflatoxins-producing fungi. Aflatoxin-degrading enzymes, which can convert poisonous aflatoxins into less dangerous compounds, are also produced by spp. The processes through which these microorganisms can be used to reduce aflatoxins in food and agricultural systems are still the subject of active research. To evaluate the novelty of tetracycline against the biosynthesis of aflatoxin in aflatoxigenic fungi via computational approach. In this study, we performed molecular docking of polyketide synthase (Pks-A), an enzyme that initiates aflatoxin biosynthesis using tetracycline, using the online SeamDock server. Our results showed that tetracycline had a strong affinity for Pks-A in the binding pocket. The binding energy of tetracycline was -12.7 kcal/mol, indicating a strong binding affinity between the two molecules. Furthermore, the binding site was located in the active site, which is a conserved region in Pks-A and is essential for catalysing the formation of aflatoxin. The results of our docking study suggest that tetracycline may be an effective inhibitor of aflatoxin biosynthesis.

Determination of aflatoxins using chromatographic methods in several foods, feed and herbal medicine products: an analytical review (from 2010 to 2022)

Aflatoxins are naturally occurring secondary metabolites generated by several molds, particularly Aspergillus flavus and Aspergillus parasiticus. They can contaminate agricultural products in the field, particularly during periods of high humidity and warmth, such as before or during harvest, or as a result of inappropriate storage, and their occurrence in the food chain is regrettably inevitable. Therefore, it is crucial to detect and measure aflatoxins. For the purpose of determining the presence of aflatoxins in various products, chromatographic methods based on high performance liquid chromatography, liquid chromatography coupled with mass spectrometry, ultra-high performance liquid chromatography coupled with mass spectrometry, high performance thin layer chromatography, and thin layer chromatography were described, along with sample preparation and risk assessment. By using several chromatographic techniques with different mobile phases and stationary phases, many analytical methods demonstrated good recovery rates of aflatoxins in a variety of products. An analysis of the existing literature indicates that from 2010 to 2022, a range of chromatographic methods have been accessible for identifying and measuring aflatoxins in food, feed, and certain herbal remedies. Although the literature shows interesting methods from an economic and environmental point of view and they can still be improved to detect aflatoxins in various products.

Risk assessments for the dietary intake aflatoxins in food: A systematic review (2016–2022)

Aflatoxins (AFs) represent one of the major food safety concerns worldwide. Agricultural commodities including cereals and their derived products, oilseeds, nuts, spices, milk and milk products are highly susceptible to aflatoxin (AF) contamination that can occur pre-harvest, during harvest or post-harvest. Aflatoxicosis is a severe health syndrome associated with direct ingestion of AFs and can be classified as acute and chronic aflatoxicosis. Complete elimination of AFs even after the food processing is hard to achieve due to their high chemical and thermal stability which further makes it extremely difficult to achieve zero exposure through the consumption of contaminated food. Risk assessments play a vital role in managing and reducing the potential risks associated with AF consumption, thereby, ensuring food and consumer safety. Therefore, a commodity wise systematic review of the risk assessments published in the last 6 years (2016–2022) has been presented in the present study in order based on the most to least no. of European Union (EU) Rapid Alert System for Food and Feed (RASFF) incidents recorded for AFs. Results suggested a significant potential health risk from the dietary exposure to AFs through the consumption of all the studied food categories (nuts, nut products and oilseeds, fruits and vegetables, herbs and spices, cereals and cereal products and milk and milk products) which held the largest number of AFs RASFF incidents. However, a need to conduct more risk assessments for ‘oilseeds’ commodity and food categories including ‘Fruits and vegetables’ and ‘Herbs and spices’ has been highlighted in the study. Additionally, a greater number of risk assessments are required to be conducted for the countries having the highest AFs RASFF notifications including Turkey, USA, Argentina, China and India. Age wise, the overall risk associated with dietary intake of AFs followed a decreasing trend with the increase in age.