Search Results (1,529)

According to the scientific information reviewed, cheese is highly susceptible to contamination by mycotoxin-producing fungi, primarily species from the genera Aspergillus (A. niger, A. flavus) and Penicillium (P. commune, P. solitum, P. palitans, and P. crustosum). Studies on various types of cheese made from cow’s milk report an average concentration of Aflatoxin M1 (AFM₁) at 13,000 ng kg⁻¹, which is alarming since the regulatory limits for AFM₁ in cheese range from 250 to 500 ng kg⁻¹. For instance, limits set by Codex Alimentarius, the European Commission (EC), Turkey, and Iran are 250 ng kg⁻¹. In the Netherlands, the limit is 200 ng kg⁻¹, and in Italy, it is 450 ng kg⁻¹. However, the concentration of mycotoxins frequently exceeds these regulatory limits, including critical mycotoxins such as ochratoxin A, citrinin, and cyclopiazonic acid, which pose significant global health concerns. Therefore, this study aims to review the mycobiota responsible for producing key mycotoxins in cheese and to assess the influence of physicochemical factors on fungal growth and mycotoxin production. By incorporating control strategies such as hygiene practices, pasteurization, and the use of preservatives, this study seeks to improve methodologies in the cheese production chain and mitigate contamination by fungi and mycotoxins. Full article

The objective of this review is to investigate the impacts of aflatoxins, particularly aflatoxin B1 (AFB₁), on intestinal microbiota, intestinal health, and growth performance in monogastric animals, primarily chickens and pigs, as well as dietary interventions to mitigate these effects. Aflatoxin B1 contamination in feeds disrupts intestinal microbiota, induces immune responses and oxidative damage, increases antioxidant activity, and impairs jejunal cell viability, barrier function, and morphology in the small intestine. These changes compromise nutrient digestion and reduce growth performance in animals. The negative impact of AFB₁ on the % change in average daily gain (ΔADG) of chickens and pigs was estimated based on meta-analysis: ΔADG (%)_chicken = −0.13 × AFB₁ intake per body weight (ng/g·d) and ΔADG (%)_pig = −0.74 × AFB₁ intake per body weight (µg/kg·d), indicating that increasing AFB₁ contamination linearly reduces the growth of animals. To mitigate the harmful impacts of AFB₁, various dietary strategies have been effective. Mycotoxin-detoxifying agents include mycotoxin-adsorbing agents, such as clay and yeast cell wall compounds, binding to AFB₁ and mycotoxin-biotransforming agents, such as specific strains of Bacillus subtilis and mycotoxin-degrading enzyme, degrading AFB₁ into non-toxic metabolites such as aflatoxin D1. Multiple mycotoxin-detoxifying agents are often combined and used together to improve the intestinal health and growth of chickens and pigs fed AFB₁-contaminated feeds. In summary, AFB₁ negatively impacts intestinal microbiota, induces immune responses and oxidative stress, disrupts intestinal morphology, and impairs nutrient digestion in the small intestine, leading to reduced growth performance. Supplementing multi-component mycotoxin-detoxifying agents in feeds could effectively adsorb and degrade AFB₁ co-contaminated with other mycotoxins prior to its absorption in the small intestine, preventing its negative impacts on the intestinal health and growth performance of chickens and pigs. Full article

The aim of this study was to estimate the aflatoxin M₁ (AFM₁) contamination in raw milk and processed milk (pasteurized or UHT) collected from two regions in Italy (Puglia and Basilicata) during a 12-year period: 2012–2023. A total of 1017 milk samples were analyzed first proceeding with screening analysis by enzyme-linked immunosorbent assay (ELISA), and suspected non-compliant samples (AFM₁ concentration higher than 0.042 µg/kg) were then analyzed by high performance liquid chromatographic with fluorimetric detection (HPLC/FLD) confirmation method. AFM₁ concentration ≥ 0.005 µg/kg (ELISA limit of quantitation) was detected in 553 of the 1017 milk samples (54.4%). AFM₁ levels exceeding the European Union maximum limit (ML) of 0.050 µg/kg were detected in 70 samples, 49 of which were determined as non-compliant samples (4.8%). Particularly high concentrations of AFM₁, exceeding 200 µg/kg, were found in four samples, three raw milk and one pasteurized. Regarding this risk exposure study, only the MOE values obtained under “high exposure scenario” were lower than 10,000, while those calculated from the overall mean values resulted as not of concern. Full article

This study aimed to explore the contamination of aflatoxins by investigating the spatial distribution of aflatoxin B1 (AFB1) in cow feedstuff and aflatoxin M1 (AFM1) in raw milk, and the potential health risks of AFM1 in milk and dairy products. Feedstuff and raw milk were collected from 160 pastures in three climate zones of China from October to November 2020. The results indicated the level of AFB1 and AFM1 ranged from 51.1 to 74.1 ng/kg and 3.0 to 7.0 ng/kg, respectively. Spatial analysis indicated the contamination was mostly concentrated in the temperate monsoon climate zone. On average, the estimated dietary exposure to AFM1 from milk and dairy products for Chinese consumers ranged from 0.0138 to 0.0281 ng/kg bw/day, with the MOE values below 10,000, and liver cancer risk of 0.00004–0.00009 cases/100,000 persons/year. For different groups, the average exposure to AFM1 was highest in the temperate monsoon climate zone and for toddlers. Full article

Cassava is an important staple food that contributes to the food security of small-scale Mozambican farmers. In southern Mozambique, cassava roots are usually processed into cassava roasted flour, locally known as “rale”. The handling and processing practices connected to “rale” production may introduce microbial contamination. We assessed the microbial contamination of “rale” processed in local farmers’ associations and consumed either locally or sold in rural markets. Microbial sampling was carried out both during the warmer rainy and cooler dry seasons, and microorganisms of relevance for food safety and fermentation were enumerated. The results revealed variation in terms of microbial diversity in all stages of cassava root processing. In samples collected in the warmer rainy season, molds, lactic acid bacteria, general aerobic bacteria and Bacillus spp. were isolated, whereas in samples collected in the cooler dry season, other groups of microorganisms such as yeasts and Staphylococcus aureus were present. Wickerhamomyces anomalus, Rhodotorula mucilaginosa, Pichia exigua, Meyerozyma caribbica and Torulaspora delbrueckii were the most frequent yeast species found within the cassava processing stages. Aflatoxin-producing molds were observed infrequently in this study, and only at low counts, thus, the risk for aflatoxin contamination appears to be low. The results obtained from the Illumina 16S rRNA gene sequencing can be considered a complementary technique to the plating methods relied on in this study. From a food quality and safety point of view, this staple food does not appear to pose a high risk for foodborne disease. Full article

Aflatoxin B1 (AFB1) contamination of food crops pose severe public health risks, particularly in decentralized agricultural systems common in low-resource settings. Effective monitoring tools are critical for mitigating exposure, but their adoption is limited by barriers such as cost, infrastructure, and technical expertise. The objectives of this study were: (1) to evaluate common AFB1 detection methods, including enzyme-linked immunosorbent assays (ELISA) and lateral-flow assays (LFA), validated via high-performance liquid chromatography (HPLC), focusing on their suitability for possible applications in decentralized, low-resource settings; and (2) to conduct a barriers-to-use assessment for commonly available AFB1 detection methods and their applicability in low-resource settings. Among four ELISA kits, the AgraQuant Aflatoxin B1 2/50 ELISA Kit demonstrated the highest accuracy and precision, reliably quantifying AFB1 in maize and tortillas across 5–150 ppb with minimal cross-reactivity. For LFA, a smartphone-based algorithm achieved a high presence/absence accuracy rate of 84% but struggled with concentration prediction. The barriers-to-use analysis highlighted the practicality of low-cost tools like moisture readers for field screening but underscored their qualitative limitations. Advanced methods like HPLC and LC-MS offer greater precision but remain impractical due to their high costs and infrastructure requirements, suggesting a potential role for adapted ELISA or LFA methods as confirmatory approaches. These findings support the development of multi-tiered frameworks integrating affordable field tools with regional or centralized confirmatory testing. Addressing systemic barriers through capacity building, partnerships, and improved logistics will enhance AFB1 monitoring in decentralized systems, protecting public health in vulnerable communities. Full article

Aspergillus flavus is a medically relevant fungus, particularly in tropical regions. Although its aflatoxin production and thermotolerance are well documented, its biofilm-forming ability has received less attention, despite being a key factor in the virulence of A. flavus as an opportunistic pathogen, which can significantly impact therapeutic outcomes. To investigate the influence of temperature on the growth and biofilm formation of an A. flavus isolate, we compared it on solid media with the reference strain A. flavus ATCC 22546 and documented morphological changes during conidial germination. We examined biofilm formation in both strains across different temperatures and evaluated the susceptibility of this A. flavus isolate to antifungal agents in both planktonic and biofilm form. Our results showed that the temperature can promote conidiation on solid media. Radial growth was highest at 28 °C, while the conidial count and density were favored at higher temperatures. Moreover, we determined that 37 °C was the optimal temperature for conidial germination and biofilm formation. We described four distinct phases in A. flavus biofilm development—initiation (0–12 h), consolidation (12–24 h), maturation (24–48 h), and dispersion (48–72 h)—with the notable presence of conidial heads at 42 °C. Carbohydrates and proteins constitute the primary components of the extracellular matrix. We observed an abundance of lipid droplets within the hyphae of the MMe18 strain biofilm. The mature biofilms demonstrated reduced susceptibility to amphotericin B and itraconazole, requiring higher inhibitory concentrations for both antifungals compared with their planktonic counterparts. Full article

It has been known since the early days of the discovery of aflatoxin B1 (AFB1) that there were large species differences in susceptibility to AFB1. It was also evident early on that AFB1 itself was not toxic but required bioactivation to a reactive form. Over the past 60 years there have been thousands of studies to delineate the role of ~10 specific biotransformation pathways of AFB1, both phase I (oxidation, reduction) and phase II (hydrolysis, conjugation, secondary oxidations, and reductions of phase I metabolites). This review provides a historical context and substantive analysis of each of these pathways as contributors to species differences in AFB1 hepatoxicity and carcinogenicity. Since the discovery of AFB1 as the toxic contaminant in groundnut meal that led to Turkey X diseases in 1960, there have been over 15,000 publications related to aflatoxins, of which nearly 8000 have addressed the significance of biotransformation (metabolism, in the older literature) of AFB1. While it is impossible to give justice to all of these studies, this review provides a historical perspective on the major discoveries related to species differences in the biotransformation of AFB1 and sets the stage for discussion of other papers in this Special Issue of the important role that AFB1 metabolites have played as biomarkers of exposure and effect in thousands of human studies on the toxic effects of aflatoxins. Dr. John Groopman has played a leading role in every step of the way—from initial laboratory studies on specific AFB1 metabolites to the application of molecular biomarkers in epidemiological studies associating dietary AFB1 exposure with liver cancer, and the design and conduct of chemoprevention clinical trials to reduce cancer risk from unavoidable aflatoxin exposures by alteration of specific AFB1 biotransformation pathways. This article is written in honor of Dr. Groopman’s many contributions in this area. Full article

Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) are considered the most important mycotoxins in terms of food safety. The aim of this study was to evaluate the hepatotoxicity of AFB1 and OTA exposure in Wistar rats and to assess the beneficial effect of fermented whey (FW) and pumpkin (P) as functional ingredients through a proteomic approach. For the experimental procedures, rats were fed AFB1 and OTA individually or in combination, with the addition of FW or a FW-P mixture during 28 days. For proteomics analysis, peptides were separated using a LC-MS/MS-QTOF system and differentially expressed proteins (DEPs) were statistically filtered (p < 0.05) distinguishing males from females. Gene ontology visualization allowed the identification of proteins involved in important biological processes such as the response to xenobiotic stimuli and liver development. Likewise, KEGG pathway analysis reported the metabolic routes as the most affected, followed by carbon metabolism and biosynthesis of amino acids. Overall, the results highlighted a strong downregulation of DEPs in the presence of AFB1 and OTA individually but not with the mixture of both, suggesting a synergistic effect. However, FW and P have helped in the mitigation of processes triggered by mycotoxins. Full article

Poultry feeds with cereal grain-based constituents are vulnerable to fungal contamination during the processing and storage stages. A total of 100 samples of compound poultry feedstuffs were collected from the cities of Riyadh, Alhassa, Qassim, and Jeddah in Saudi Arabia. A quantitative enumeration of fungal colony-forming units (CFUs) was performed on Dichloran Rose Bengal Chloramphenicol Agar (DRBC) and Czapek Iprodione Dichloran Agar (CZID) media. Aspergillus flavus was the most predominant species, accounting for 18.714 × 10³ and 3.956 × 10³ CFU/g, with frequencies of 84 and 42% in the feed samples on DRBC and CZID media, respectively. The levels of different mycotoxins were estimated by the HPLC technique. One hundred percent of the compound poultry feedstuff samples were contaminated by mycotoxins such as AFB1, AFB2, AFG1, AFG2, FB1, DON, T2, OTA, and ZEN. Aflatoxins were recorded in 84% of the tested samples, of which 70 samples were contaminated by AFB1, ranging from 0.03 to 0.40 μg/kg. The aflatoxin analysis of the fungal species revealed that 89% and 100% of A. flavus and A. parasiticus isolates were aflatoxigenic, and all of them exhibited the presence of the aflR, omt-1, ver-1, and nor-1 genes. According to the PCR protocol based on FLA1, two primers were successful in directly and rapidly detecting A. flavus in the poultry feedstuff samples. Full article

Background/Objectives: Mycotoxins, secondary metabolites synthesized by filamentous fungi, have been classified as dangerous substances and proven to be carcinogenic, as well as to have genotoxic, nephrotoxic, hepatotoxic, teratogenic, and mutagenic properties. Despite numerous trials to develop an effective and safe-for-human-health method of detoxification, there is still a high risk associated with the occurrence of these toxins in food and feed. Biological methods of food preservation are an alternative option to conventional chemical and physical methods, characterized by their less negative impact on human health as well as their high efficiency against filamentous fungi and other foodborne pathogens. Mycoremediation is a new biotechnique based on the capability of fungi to detoxify matrices from various pullulans. Ligninolytic enzymes produced by white rot fungi (WRF) characterize a high efficiency in the degradation of various mycotoxins. Methods: In our study, Pleurotus ostreatus, as a representative of WRF, was cultivated on a medium contaminated by AFB1 and ZEN (mushroom substrate and maize) in a few variants of concentration. After the cultivation, medium and fruiting bodies were collected and analyzed with the usage of HPLC and LC/MS methods. Results: The reduction oscillated between 53 and 87% (AFB1) and 73 and 97% (ZEN) depending on the initial concentration of toxins in the medium. Grown fruiting bodies contained insignificant amounts of both toxins. Conclusions: These findings confirm the potential of P. ostreatus as an effective biological agent for reducing mycotoxins in contaminated medium, highlighting its applicability in developing sustainable and safe methods for detoxification. Full article

Mycotoxins are secondary metabolites of filamentous fungi that cause massive agricultural losses worldwide and constitute a significant health problem for humans and animals. The aim of this five-year study was to investigate the contamination of compound feed for broiler chickens at all stages (starter, grower and finisher) from a feed mill in Romania with mycotoxins such as total aflatoxins (AFT), deoxynivalenol (DON), fumonisins (FUMs), ochratoxin A (OTA) and zearalenone (ZEN). AFT was detected in 49.3–72.2% of the samples with concentrations ranging from 0.01 to 5.2 µg/kg. DON was detected in 77.6–98.9% of the samples, with maximum concentrations ranging from 330 to 1740 µg/kg. FUM contamination ranged from 42.7% to 87.2%, with maximum levels between 460 and 1400 µg/kg. OTA was present in 44.2–87.9% of the samples, with maximum concentrations reaching 21.4 µg/kg. ZEN was consistently elevated at all feeding stages, being detected in 86.5–97.4% of the samples, with maximum levels of 89.4 µg/kg. Mycotoxin co-occurrence was common in the samples, with the most common combination of four mycotoxins occurring in 38.51% of the samples. Samples were collected from storage bunkers, homogenised and analysed in certified laboratories, with sampling procedures varying according to batch size to ensure representativeness. Investigation of the transfer of mycotoxins into animal products and the combined effects of mycotoxins on animal health, including potential synergistic or antagonistic interactions, is particularly relevant. This study emphasises the essential role of comprehensive and continuous monitoring of mycotoxins in protecting animal health and food safety. Full article

Background: The Mycotoxin Mitigation Trial (MMT) was a community-based cluster-randomized trial designed to assess the effect of dietary aflatoxin (AF) on linear growth. Similar dietary intake between arms was an important component of the trial’s program theory and essential for the trial’s internal validity and interpretation. Objective: This analysis assessed and compared dietary intake by arm within a sub-sample of infants enrolled in the MMT. Methods: Twenty paired clusters (10 per trial arm) out of the 52 MMT clusters were included in this sub-sample. Up to 15 maternal/infant dyads per cluster were randomly selected for a one-time, structured, multi-pass 24 h dietary recall. Data were collected at the midpoint of the trial, when infants were 12 months of age, over 8 calendar months. We evaluated and compared infant nutrient intake and adequacy of energy, protein, lipid, iron, zinc, calcium, and vitamin A between study arms. Nutrient intake by arm was estimated using mixed-level regression models. Results: A total of 282 mothers participated (n = 140 intervention arm and 142 standard of care (SoC) arm). The mean daily intakes of energy and lipid fed to infants were 505 kcal/day (SD = 225.9) and 13 g/day (SD = 6.9), respectively, in the intervention and SoC arms, with no difference between arms. Intervention infants consumed slightly more protein than SoC infants (13.7 v. 12.3 g/day, p = 0.02). Consumption of iron, zinc, calcium, and vitamin A were low and did not differ between arms. Conclusions: At the midpoint of the MMT, energy, lipid, and micronutrient intake did not differ between arms. Protein consumption was slightly greater in the intervention arm. Guided by the trial’s program theory, this analysis advances the interpretation of the MMT trial findings. Full article

Maize is one of the major crops that are susceptible to Aspergillus flavus infection and subsequent aflatoxin contamination, which poses a serious health threat to humans and domestic animals. Here, an RNA interference (RNAi) approach called Host-Induced Gene Silencing (HIGS) was employed to suppress the O-methyl transferase gene (omtA, also called aflP), a key gene involved in aflatoxin biosynthesis. An RNAi vector carrying part of the omtA gene was introduced into the B104 maize line. Among the six transformation events that were positive for containing the omtA transgene, OmtA-6 and OmtA-10 were self-pollinated from T1 to T4, and OmtA-7 and OmtA-12 to the T6 generation. These four lines showed at least an 81.3% reduction in aflatoxin accumulation at the T3 generation under laboratory conditions. When screened under field conditions with artificial inoculation, OmtA-7 at T5 and T6 generations and OmtA-10 at T4 generation showed a reduction in aflatoxin contamination between 60% and 91% (p < 0.02 to p < 0.002). In order to develop commercial maize lines with enhanced aflatoxin resistance, the omtA transgene in OmtA-7 was introduced into three elite inbred lines through crossing, and the resulting crosses also exhibited significantly lower aflatoxin accumulation compared to crosses with non-transgenic controls (p < 0.04). In addition, high levels of omtA-specific small RNAs were only detected in the transgenic kernel and leaf tissues. These results demonstrate that suppression of omtA through HIGS can enhance maize resistance to aflatoxin contamination, and this resistance can be transferred to elite backgrounds, providing a viable and practical approach to reduce aflatoxin contamination in maize. Full article

Aflatoxins are foodborne toxins that occur naturally in various crops because of fungal contamination, particularly from two strains, namely Aspergillus flavus and Aspergillus parasiticus. Given their adverse properties, which are teratogenic, mutagenic, and carcinogenic, aflatoxins present a significant public health concern. Consequently, efforts are underway to inactivate aflatoxins and inhibit the growth of these fungi to prevent toxin formation. Since chemical treatments for food products are undesirable or even restricted in some countries, alternative approaches are also implemented. This study investigated gamma-ray (γ-ray) irradiation as a potential method for reducing aflatoxin levels. Specifically, solutions of aflatoxins B1, B2, G1, and G2 were irradiated with doses of 1, 2, 4, and 8 kGy using a cobalt-60 irradiation source. Following γ-irradiation, a notable reduction in aflatoxin levels was observed, particularly for types B1 and G1, which process higher toxicity. This finding suggests γ-irradiation as a feasible method for aflatoxin deactivation. Additionally, as a proof of concept, almond samples spiked with aflatoxins and A. flavus were irradiated. The results showed a decrease in both aflatoxin levels and microbial load in these samples. Overall, these findings indicate that γ-irradiation is a promising approach to aflatoxin reduction, microbial decontamination, and the potential extension of almonds’ shelf life. Full article