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Tobacco contains many harmful chemicals, toxins, and carcinogens which cause serious health hazards. Mycotoxins and Aflatoxins are present in all tobacco including chewing and smokeless tobacco. We detected and isolated the aflatoxins-producing fungi Aspergillus sp. by microbial and molecular-based techniques. 50 samples were collected randomly and grown on SDA media by spread plate method. Isolates of A. flavus were analyzed for Aflatoxin production by Ammonia vapor test and the DNA of isolates was extracted and subjected for further molecular identification. Different fungal genera were observed including the most common Aspergillus while the other were Penicillium, Rhizopus, candida, and Trichophyton. Aspergillus flavus was isolated and characterized as a white soft velvety surface. The diameter of colonies was observed as 50 - 70 mm. Coconut-based medium detection by UV light of 365 nm showed the production of blue-green fluorescence on the reverse of colonies on coconut agar medium. DNA extracted from all the strains producing aflatoxins was quantified by VICAM. High levels of Aflatoxin ranging from 21 - 47 ug/kg were detected in 10 different samples while the remaining samples had a range of 10 - 20 ug/kg. the study showed both quantitative and qualitative techniques for the detection of aflatoxin-producing fungi in chewing tobacco. aflR and aflS were observed as potential candidates for the detection of aflatoxigenic fungi. [ABSTRACT FROM AUTHOR]

Aspergillus flavus is a major aflatoxin B1, posing significant health concerns to humans, crops, and producer fungi. Due to the undesirable consequences of the usage of synthetic fungicides, biological control using yeasts has gained more attention. In this study, eight isolates of epiphytic yeasts belonging to Moesziomyces sp., Meyerozyma sp. and Metschnikowia sp., which have been identified as antagonists, were isolated from different plants, including grapes, blueberries, hawthorns, hoşkıran, beans and grape leaf. While volatile organic compounds (VOCs) produced by Moesziomyces bullatus DN-FY, Metschnikowia aff. pulcherrima DN-MP and Metschnikowia aff. pulcherrima 32-AMM reduced in vitro A. flavus mycelial growth and sporulation, only VOCs produced by Metschnikowia aff. fructicola 1-UDM were found to be effective at reducing in vitro AFB1 production. All yeasts reduced the mycelial growth of A. flavus by 76–91%, while AFB1 production reduced to 1.26–10.15 ng/g and the control plates' growth was 1773 ng/g. The most effective yeast, Metschnikowia aff. Pulcherrima DN-HS, reduced Aspergillus flavus growth and aflatoxin B1 production on hazelnuts. The AFB1 content on hazelnuts reduced to 333.01 ng/g from 536.74 ng/g. To our knowledge, this is the first report of yeasts isolated from plants being tested as potential biological control agents to reduce AFB1 production on hazelnuts. [ABSTRACT FROM AUTHOR]

Aflatoxins are among the main carcinogens threatening food and feed safety while imposing major detection challenges to the agrifood industry. Today, aflatoxins are typically detected using destructive and sample-based chemical analysis that are not optimally suited to sense their local presence in the food chain. Therefore, we pursued the development of a non-destructive optical sensing technique based on fluorescence spectroscopy. We present a novel compact fluorescence sensing unit, comprising both ultraviolet excitation and fluorescence detection in a single handheld device. First, the sensing unit was benchmarked against a validated research-grade fluorescence setup and demonstrated high sensitivity by spectrally separating contaminated maize powder samples with aflatoxin concentrations of 6.6 µg/kg and 11.6 µg/kg. Next, we successfully classified a batch of naturally contaminated maize kernels within three subsamples showing a total aflatoxin concentration of 0 µg/kg, 0.6 µg/kg and 1647.8 µg/kg. Consequently, our novel sensing methodology presents good sensitivity and high potential for integration along the food chain, paving the way toward improved food safety. [ABSTRACT FROM AUTHOR]

The SARS-CoV-2 virus caused a major transformation in the food system globally including in Rwanda. This research identified and assessed the status and structure of the maize production system in the Kigali city region before and during the COVID-19 pandemic. The methods adopted for this study include both quantitative and qualitative methods using primary data obtained from the participants' interview and focus group discussions, the secondary data were obtained from national institute of statistics of Rwanda (NISR). While production, processing, distribution and consumption are the four parts of the maize production system, the study only focused on the production system. The sample size for the study was 256 respondents who were maize production system actors from the Kigali city region. The study showed that before the COVID-19 pandemic, the prize of dried maize was significantly influenced by the cost of diammonium phosphate (DAP) (p=0.000), the source of the irrigating scheme (p=0.008), being a cooperative member (p=0.000) and marital status (p=0.002). During the pandemic, DAP (p=0.109) was absent at market due to lockdowns, and farmers did not access it. Maize farmers-built responses of resilience, persistence, adaptation, transformation, and persistence to encounter the consequences of the lockdowns. Innovative responses to shortage of maize- input stocks, and poor imports were presented, which represented 26.66% of resilient responses adopted by farmers. The innovative responses to labor shortage with farm workers' migration to their home provinces before and during the implementation of containment measures were 20%. Resilience built against the absence of extension services due to restricted movements was 13.33%. Reactions to restricted movement to and from fields, and adaptation to COVID-19 pandemic containing measures were 26.66 %, while reaction to the shortage of dried -maize at the market, which shortened the maize production cycle was 13.33%. Apart from maize production, more than half of the maize selling system were women 51.61%. Women in raw maize processing were 67.67% with a 100% level of university. [ABSTRACT FROM AUTHOR]

This study reports onthe effect of various production steps on levels of aflatoxins during preparation of thobwa, a traditional maize‐based fermented non‐alcoholic beverage, brewed across Malawi. The effect of boiling, fermentation and their interaction on the level of aflatoxins, the trends of aflatoxin reduction during brewing, and the distribution of aflatoxins between the solid and liquid phases of the beverage were studied using VICAM AflaTest immunoaffinity fluorometric assay. Fermenting and boiling thobwa pre‐mix‐, with initial aflatoxin content of 45–183 μg/kg, resulted in aflatoxin reduction of 47% (13–61 μg/kg) on average. Fermentation and boiling contributed about 20 and 33% aflatoxin reduction, respectively, but without interactive effect between the two factors. Fermenting the thobwa for 24 h led to further reduction of aflatoxins to about 37% of the initial content, and remainedconstant for up to 8 days. Thobwa is a popular beverage in Malawi which is consumed in large quantities by all gender categories including infants, therefore the presence of aflatoxins may constitute a significant health risk factor. This study highlights the need to use raw materials with low levels of aflatoxins for production of maize‐based non‐alcoholic beverages to ensure consumer safety. [ABSTRACT FROM AUTHOR]

The preharvest maize mycobiome may be crucial in defining the health of the crop in terms of potential disease burden and mycotoxins. We investigated the preharvest maize mycobiome structure, including the influence of weather patterns, in terms of rainfall intensity, on its composition. In addition, we investigated correlation of genera Fusarium and Aspergillus with maize fumonisin-B1 and aflatoxin. Forty maize fields from selected districts in the wetter northern (N) and drier southern (S) agroecological zones of Zambia were sampled twice over two seasons (1 and 2). The defined weather variables over the two seasons were low rainfall with dry spell (S1), low rainfall (S2), and high rainfall (N1 and N2). High-throughput DNA amplicon sequencing of internal transcribed spacer 1 (ITS1) was used to determine the mycobiome structure and the composition in relation to rainfall patterns. We detected 61 genera, with Fusarium and previously unreported Sarocladium in Zambia to have the highest frequency of detection on the maize. There was a significant difference in fungal genera composition between S1 and S2 but no difference between N1 and N2. The weather pattern with dry spell, S1, had a strong proliferation of Meyerozyma and xerophiles Penicillium, Kodamaea, and Aspergillus. The four genera drove the difference in composition between S1 and S2 and the significantly higher fungal diversity in S1 compared to N2. Of the mycotoxin-important fungi, dry conditions (S1) were a key driver for proliferation of Aspergillus, while Fusarium proliferation occurred irrespective of weather patterns. The relative abundance of Aspergillus and Fusarium resonated with maize aflatoxin and fumonisin-B1 levels, respectively. IMPORTANCE Fungi contaminate various crops worldwide. Maize, an important human staple and livestock cereal, is susceptible to contamination with fungi in the field. Fungi are drivers of plant disease and can compromise yield. Some species of fungi are known to produce chemical compounds (mycotoxins), which are cancer-causing agents in humans and impair livestock productivity. It is important to understand the spectrum of fungi on maize and how weather conditions can impact their abundance. This is because the abundance of fungi in the field can have a bearing on the health of the crop as well as potential for mycotoxins contamination. By understanding the spectrum of the preharvest fungi, it becomes possible to know the key fungi adapted to the maize and subsequently the potential for crop disease as well as mycotoxins contamination. The influence of weather conditions on the spectrum of preharvest fungi on maize has not been fully explored. [ABSTRACT FROM AUTHOR]

In this study, the average level of aflatoxin M1 in various types of milk from 107 articles (297 studies with 16,274 milk samples) were meta-analyzed using random-effect model based on the milk varieties (animal species and heating processes), geographical regions, seasons, detection techniques and dairy farming subgroups. Studies on milk contamination with aflatoxin M1 in Iran were collected using universal and Persian databanks from January 1974 to the end of November 2021. The overall aflatoxin M1 mean concentration and prevalence in milk samples of Iran were 39.65 ng/l (95% CI: 36.00–43.30) and 80% (95% CI: 76–85%), respectively. The rank order of importance of various variables in mean levels of aflatoxin M1 in milk samples included milk type (animal species) > geographical regions > detection techniques > dairy farming types > milk types (heating processes) > seasons. Findings revealed that the overall content of aflatoxin M1 in milk samples of Iran was lower than that allowed by the European Union, Institute of Standards and Industrial Research of Iran, and the USA, possibly due to the milk monitoring by the Iranian regulatory systems. [ABSTRACT FROM AUTHOR]

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The apparent rarity of sex in many fungal species has raised questions about how much sex is needed to purge deleterious mutations and how differences in frequency of sex impact fungal evolution. We sought to determine how differences in the extent of recombination between populations of Aspergillus flavus impact the evolution of genes associated with the synthesis of aflatoxin, a notoriously potent carcinogen. We sequenced the genomes of, and quantified aflatoxin production in, 94 isolates of A. flavus sampled from seven states in eastern and central latitudinal transects of the United States. The overall population is subdivided into three genetically differentiated populations (A, B, and C) that differ greatly in their extent of recombination, diversity, and aflatoxin-producing ability. Estimates of the number of recombination events and linkage disequilibrium decay suggest relatively frequent sex only in population A. Population B is sympatric with population A but produces significantly less aflatoxin and is the only population where the inability of nonaflatoxigenic isolates to produce aflatoxin was explained by multiple gene deletions. Population expansion evident in population B suggests a recent introduction or range expansion. Population C is largely nonaflatoxigenic and restricted mainly to northern sampling locations through restricted migration and/or selection. Despite differences in the number and type of mutations in the aflatoxin gene cluster, codon optimization and site frequency differences in synonymous and nonsynonymous mutations suggest that low levels of recombination in some A. flavus populations are sufficient to purge deleterious mutations.IMPORTANCE Differences in the relative frequencies of sexual and asexual reproduction have profound implications for the accumulation of deleterious mutations (Muller's ratchet), but little is known about how these differences impact the evolution of ecologically important phenotypes. Aspergillus flavus is the main producer of aflatoxin, a notoriously potent carcinogen that often contaminates food. We investigated if differences in the levels of production of aflatoxin by A. flavus could be explained by the accumulation of deleterious mutations due to a lack of recombination. Despite differences in the extent of recombination, variation in aflatoxin production is better explained by the demography and history of specific populations and may suggest important differences in the ecological roles of aflatoxin among populations. Furthermore, the association of aflatoxin production and populations provides a means of predicting the risk of aflatoxin contamination by determining the frequencies of isolates from low- and high-production populations.

Pre-mRNA encoding human NEIL1 undergoes editing by adenosine deaminase ADAR1 that converts a single adenosine to inosine, and this conversion results in an amino acid change of lysine 242 to arginine. Previous investigations of the catalytic efficiencies of the two forms of the enzyme revealed differential release of thymine glycol (ThyGly) from synthetic oligodeoxynucleotides, with the unedited form, NEIL1 K242 being ≈30-fold more efficient than the edited NEIL1 K242R. In contrast, when these enzymes were reacted with oligodeoxynucleotides containing guanidinohydantoin or spiroiminohydantoin, the edited K242R form was ≈3-fold more efficient than the unedited NEIL1. However, no prior studies have investigated the efficiencies of these two forms of NEIL1 on either high-molecular weight DNA containing multiple oxidatively-induced base damages, or oligodeoxynucleotides containing a bulky alkylated formamidopyrimidine. To understand the extent of changes in substrate recognition, γ-irradiated calf thymus DNA was treated with either edited or unedited NEIL1 and the released DNA base lesions analyzed by gas chromatography-tandem mass spectrometry. Of all the measured DNA lesions, imidazole ring-opened 4,6-diamino-5-formamidopyrimidine (FapyAde) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) were preferentially released by both NEIL1 enzymes with K242R being ≈1.3 and 1.2-fold more efficient than K242 on excision of FapyAde and FapyGua, respectively. Consistent with the prior literature, large differences (≈7.5 to 12-fold) were measured in the excision of ThyGly from genomic DNA by the unedited versus edited NEIL1. In contrast, the edited NEIL1 was more efficient (≈3 to 5-fold) on release of 5-hydroxycytosine. Excision kinetics on DNA containing a site-specific aflatoxin B1-FapyGua adduct revealed an ≈1.4-fold higher rate by the unedited NEIL1. Molecular modeling provides insight into these differential substrate specificities. The results of this study and in particular, the comparison of substrate specificities of unedited and edited NEIL1 using biologically and clinically important base lesions, are critical for defining its role in preservation of genomic integrity.

Almond is a desirable and high-quality food source where the presence of nut allergens and a vulnerability to aflatoxin and Salmonella contamination represent threats to consumer safety. In 2019, over 1 billion kg. of almonds, representing over 80% of the world total, were produced in California from a relatively few varieties with a very narrow genetic base. To address emerging needs mandated by cultural and climate changes, new germplasm has been introduced combining peach as well as wild peach and wild almond species. Advanced breeding selections incorporating exotic germplasm into a genetic background compatible with commercial production in California have demonstrated sizable reductions in level of kernel immunoreactivity as well as opportunities for improved control of aflatoxin and Salmonella. Breeding strategies employed include direct selection for reduced kernel immunoreactivity from an introgression enriched germplasm, the integration and pyramiding of resistance to multiple components of the aflatoxin disease-insect complex, and introduction of novel nut and tree traits to facilitate mechanized catch-frame field harvesting to avoid contamination with soil-borne pathogens such as Salmonella and Escherichia coli, as well as agrochemical residues.

The cultivation and the usage of maize have been significantly increased across all provinces of Rwanda. Nevertheless, the problem of aflatoxin contamination remains a major factor that renders them to be unfit for animal and human consumption. In this research, the effects of drying methods (sun drying, kitchen drying, dry shelter) post aflatoxin infection in maize were evaluated in Northern Rwanda. A randomized complete block design with four replications was used with maize variety (H628) with drying methods as the main plot. Maize samples were tested for aflatoxin using aflatest. In both seasons (2021 A and 2021 B) the aflatoxin results show that the aflatoxin infection levels were lower in maize dried with kitchen drying (1.4 ppb) compared to the samples dried with sun drying (1.6 ppb) and dry shelter (2.2). Also, it was seen that the three drying methods are good for reducing the aflatoxin infection levels lower than 10 ppb as standards limit for East African countries. The aflatoxin infection levels were found to be significant (≤10 ppb) as results of kitchen drying than sun drying and dry shelter methods. This research resulted that proper drying such as drying maize on kitchen fire (kitchen drying) produce the lower level of aflatoxin infection in maize.

Background & Aims: Aflatoxin exposure increases the risk for hepatocellular carcinoma (HCC) in hepatitis B virus (HBV)-infected individuals, particularly males. We investigated sex-based differences in the HCC genome and antitumor immunity. Methods: Whole-genome, whole-exome, and RNA sequencing were performed on 101 HCC patient samples (47 males, 54 females) that resulted from HBV infection and aflatoxin exposure from Qidong. Androgen on the expression of aflatoxin metabolism-related genes and nonhomologous DNA end joining (NHEJ) factors were examined in HBV-positive HCC cell lines, and further tested in tumor-bearing syngeneic mice. Results: Qidong HCC differed between males and females in genomic landscape and transcriptional dysfunction pathways. Compared with females, males expressed higher levels of aflatoxin metabolism-related genes, such as AHR and CYP1A1, and lower levels of NHEJ factors, such as XRCC4, LIG4, and MRE11, showed a signature of up-regulated type I interferon signaling/response and repressed antitumor immunity. Treatment with AFB1 in HBV-positive cells, the addition of 2 nmol/L testosterone to cultures significantly increased the expression of aflatoxin metabolism-related genes, but reduced NHEJ factors, resulting in more nuclear DNA leakage into cytosol to activate cGAS-STING. In syngeneic tumor-bearing mice that were administrated tamoxifen daily via oral gavage, favorable androgen signaling repressed NHEJ factor expression and activated cGAS-STING in tumors, increasing T-cell infiltration and improving anti–programmed cell death protein 1 treatment effect. Conclusions: Androgen signaling in the context of genotoxic stress repressed DNA damage repair. The alteration caused more nuclear DNA leakage into cytosol to activate the cGAS-STING pathway, which increased T-cell infiltration into tumor mass and improved anti–programmed cell death protein 1 immunotherapy in HCCs.

BackgroundAspergillus spp. comprises a very diverse group of lower eukaryotes with a high relevance for industrial applications and clinical implications. These multinucleate species are often cultured for many generations in the laboratory, which can unknowingly propagate hidden genetic mutations. To assess the likelihood of such events, we studied the genome stability of aspergilli by using a combination of mutation accumulation (MA) lines and whole genome sequencing.ResultsWe sequenced the whole genomes of 30 asexual and 10 sexual MA lines of three Aspergillus species (A. flavus, A. fumigatus and A. nidulans) and estimated that each MA line accumulated mutations for over 4000 mitoses during asexual cycles. We estimated mutation rates of 4.2 × 10-11 (A. flavus), 1.1 × 10-11 (A. fumigatus) and 4.1 × 10-11 (A. nidulans) per site per mitosis, suggesting that the genomes are very robust. Unexpectedly, we found a very high rate of GC → TA transversions only in A. flavus. In parallel, 30 asexual lines of the non-homologous end-joining (NHEJ) mutants of the three species were also allowed to accumulate mutations for the same number of mitoses. Sequencing of these NHEJ MA lines gave an estimated mutation rate of 5.1 × 10-11 (A. flavus), 2.2 × 10-11 (A. fumigatus) and 4.5 × 10-11 (A. nidulans) per base per mitosis, which is slightly higher than in the wild-type strains and some ~ 5-6 times lower than in the yeasts. Additionally, in A. nidulans, we found a NHEJ-dependent interference of the sexual cycle that is independent of the accumulation of mutations.ConclusionsWe present for the first time direct counts of the mutation rate of filamentous fungal species and find that Aspergillus genomes are very robust. Deletion of the NHEJ machinery results in a slight increase in the mutation rate, but at a rate we suggest is still safe to use for biotechnology purposes. Unexpectedly, we found GC→TA transversions predominated only in the species A. flavus, which could be generated by the hepatocarcinogen secondary metabolite aflatoxin. Lastly, a strong effect of the NHEJ mutation in self-crossing was observed and an increase in the mutations of the asexual lines was quantified.

ObjectiveContamination of pistachio with Aspergillus and aflatoxin is one of the most important problems in the production and export of this valuable product. In this research, in addition to molecular identification of Aspergillus flavus and A. parasiticus and determination of the frequency of their toxigenic and atoxigenic isolates, the population of Aspergillus flavus clade was measured in soil, intact and cracked fruits of cv. Ohadi in 6 treatments of last irrigation before harvest (5, 10, 15, 20, 25 and 30 days). Materials and methodsIsolation of Aspergillus flavus clade fungi from soil, intact and cracked fruits was done by serial dilution method on DRBC culture medium. Aspergillus flavus and A. parasiticus were identified by macromorphological and molecular (calmodulin primer) features and screening of toxigenic and atoxigenic isolates was performed using of coconut-agar medium (CAM) and thin layer chromatography (TLC). This experiment was carried out in a complete randomized block design with four replications during the years 2012 to 2014. ResultsOut of 233 collected isolates of Aspergillus flavus clade, 221 and 12 isolates belonged to Aspergillus flavus and A. parasiticus, respectively. Frequency of toxigenic isolates ofA. flavus and A. parasiticus was 86.5, 87.5, 88.7% and 80, 75, 100% in cracked, intact fruits and soil, respectively. Based on the results of the combine analysis, the last irrigation treatments 30 and 25 days before harvest have the lowest and the last irrigation treatments 10 and 5 days before harvest have the highest population of Aspergillus flavus clade in the soil, intact and cracked fruits which showed significant difference with each other. Using of manure significantly increased the population of Aspergillus flavus clade, so that this increase in cracked, healthy fruits and soil in 2013 was 66, 83 and 114%, respectively. ConclusionsThe results of the present research showed that reduction of the interval between the last irrigation and the time of fruit harvesting (high moisture percentage of the surface soil of the orchard) and the using manure can increase the population of Aspergillus flavus clade in the soil, intact and cracked pistachio fruits. Increasing of the fungal population is accompanied by the frequency of toxigenic isolates of Aspergillus flavus and A. parasiticus. Adjusting the irrigation cycle, so that the last irrigation is 25 to 30 days before the fruit harvesting, can reduce the possibility of contamination of the pistachio kernels to Aspergillus and aflatoxin.

The objective of this review paper was to assess the effect of aflatoxin contamination in dairy products and its toxicity on public health in the Ethiopian dairy sector. Feed contamination by mycotoxins causes serious issues with the economy, food security, and safety. The economic impact can take different forms, direct market costs of missed trade or lower profits resulting from the rejection of contaminated animal products, reduced productivity, animal death, particularly in more sensitive calves, and increased treatment. Nougcake, which is often used as feed for dairy animals, is a significant source of aflatoxin contamination in the peri-urban dairy value chain in Addis Ababa. Aflatoxin AFM is the primary hydroxylated AFB metabolite in milk from cows that were discharged after consuming a meal contaminated with AFB for 12 hours. Prolonged or chronic exposure to aflatoxins has a number of harmful effects on health, including potent carcinogens and may affect all organ systems, especially the liver and kidneys; that cause liver cancer, mutagenic, hepatotoxic, carcinogenic, and teratogenic effects on cattle. According to Brazilian law, the maximum permitted levels for liquid milk, milk powder, and cheese are 0.5mg/kg,5.0mg/kg and 2.5mg/kg, respectively. As a result, there is increased aflatoxin contamination in both humans and animals. In conclusion, as compared to other countries, the effect of aflatoxin contamination and its toxicity was higher in Ethiopian dairy sector. In order to regulate aflatoxin contamination all coordinated efforts from all relevant groups should work together and further intervention should be implemented via policymakers, dairy sectors, Government and non-Governmental organizations.

Mycotoxins can accumulate in raw materials of plant origin at different technological stages of its production. Most often, the producers of mycotoxins are fungi of the genera Aspergillus , Fusarium , Penicillium and some others. The clinical symptoms of mycotoxicoses vary significantly, and lethal outcomes are possible. For this reason, the mycotoxicological study of various types of feed under production conditions is an indispensable component of veterinary support of industrial animal husbandry. As part of this study, a retrospective analysis of the results of a mycotoxicological study of feed for different animal species was carried out in the Omsk region in 2017-2021. All feeds received by the Omsk Regional Veterinary Laboratory for 5 years for the determination of mycotoxins were examined for the presence of ochratoxin A, zearalenone, T-2 toxin, aflatoxin B 1, deoxynivalenol. It was established that almost 70 % of the studied samples contained mycotoxins, including their maximum allowable level was exceeded in 74 samples. Exceeding the permissible levels was noted for the content of T-2 toxin (34 samples), zearalenone (27 samples), ochratoxin A (6 samples), aflatoxin B 1 (4 samples) and deoxynivalenol (3 samples). The largest number of cases of contamination was recorded in the study of feed and feed mixtures. The greatest danger is the multiple contamination of feed with mycotoxins. This increases the risk of developing comorbid conditions and the spread of opportunistic infections.

Aflatoxin M1 was determined in 240 milk samples collected from Hulla and Dale districts, Ethiopia, by Enzyme linked immunosorbent assays (ELISA) detection. It has been found that 96.7% of the respondents did not have any awareness about aflatoxin. Laboratory analysis showed that AFM1 was detected in 77.91% of the total raw milk samples collected within a range of 0.00–0.038 and with an average of 0.0068 µg/L. Average AFM1 in raw milk during dry season (0.008 ± 0.0093 µg/L) was significantly higher (P

Aflatoxin contamination in food commodities is a major food safety concern all over the world. These aflatoxins are a group of carcinogenic secondary metabolites produced by some fungi belonging to Aspergillus section Flavi. To minimize the level of aflatoxins in food commodities, a wide range of pre- and post-harvest procedures have been employed. In this study, aqueous extracts of 13 medicinal herbs were assessed for their ability to detoxify aflatoxin B1 (AFB1), the most potent and commonly occurring carcinogen in foods. Among them, herbal extracts of Hybanthus enneaspermus, Eclipta prostrata and Centella asiatica exhibited over 70% detoxification of AFB1. The degradation of AFB1 upon treatment with these herbal extracts was confirmed by liquid chromatography–mass spectrometry analysis. Two fractions (Rf 0.75 and 0.87) purified from H. enneaspermus by using thin-layer chromatography displayed in vitro AFB1-detoxifying properties. Gas chromatography–mass spectrometry analysis of the active fractions revealed the presence of linalool and bornyl acetate as the major components suggesting possible involvement of volatile compounds of H. enneaspermus in the detoxification of AFB1. To our knowledge, this is the first report on the detoxification of AFB1 by H. enneaspermus, E. prostrata and C. asiatica aqueous extracts.

为了开发适用于食用油的安全、高效黄曲霉毒素吸附剂，以一种半干法制备的钙基蒙脱土为研究对象，采用单因素实验和正交实验优化了蒙脱土脱除花生油中黄曲霉毒素B1的工艺,利用等温吸附实验研究了其吸附机制，并考察蒙脱土吸附对花生油品质的影响。结果表明：在蒙脱土添加量0.5%、吸附温度110 ℃、吸附时间30 min的条件下，黄曲霉毒素B1的脱除率最高，为（95.15±0.01）%，花生油中黄曲霉毒素B1的含量降低至（1.995±0.607） μg/kg，符合国家标准（GB 2761—2017）要求；蒙脱土吸附花生油中黄曲霉毒素B1符合Ferundlich模型，表明该吸附是物理多层吸附和化学单层吸附的复杂过程；蒙脱土对黄曲霉毒素B1的吸附亲和力强，最大吸附量可达0.1 mg/g；该蒙脱土还可改善花生油的色泽，降低花生油的酸值和过氧化值，且总生育酚和总甾醇保留率较高。该蒙脱土是一种安全、高效的黄曲霉毒素B1吸附剂，可用于油脂加工中。 To develop a safe and high-efficiency adsorbent for removal afloxins from edible oil, a calcium-based montmorillonite prepared by semi-dry method was used as a research object. The removal process of aflatoxin B1 (AFB1) from peanut oil by montmorillonite was optimized by single factor experiment and orthogonal experiment and the adsorption mechanism was studied by isothermal adsorption experiment. In addition, the effect of montmorillonite on the peanut oil quality was studied. The results showed that under the conditions of montmorillonite dosage 0.5%, adsorption temperature 110 ℃ and adsorption time 30 min, the removal rate of AFB1 was the highest, which was (95.15±0.01)%, and the content of AFB1 in peanut oil was (1.995±0.607) μg/kg, which could meet the national standard (GB 2761-2017). The adsorption process of AFB1 in peanut oil by montmorillonite could be well fitted with Freundlich equation, indicating the adsorption of AFB1 by montmorillonite was a complex process including physical multilayer adsorption and chemical monolayer adsorption. The montmorillonite had a strong adsorption affinity for AFB1 and the highest adsorbing capacity was up to 0.1 mg/g. The montmorillonite also could improve the color, and reduce the acid value and peroxide value of peanut oil. The retention rates of total sterols and total tocopherols were high. The montmorillonite is a safe and efficient AFB1 adsorbent, and can be used in oil processing.

AbstractFilamentous fungi that could be classified into Aspergillus flavus/oryzae were isolated from traditionally fermented meju commercially available in Korea. The samples were analyzed for aflatoxin B1 and ochratoxin A contamination by HPLC; however, no toxin was detected. In addition, fungal and bacterial metagenomic sequencing were performed to analyze the microbial distribution in the samples. The results revealed that the distribution and abundance of fungi and bacteria differed considerably depending on the production regions and fermentation conditions of the meju samples. Through morphological analysis, ITS region sequencing, and assessment of the aflatoxin-producing ability, a total of 32 A. flavus/oryzae strains were identified. PCR analysis of six regions with a high mutation frequency in the aflatoxin gene cluster (AGC) revealed a total of six types of AGC breaking point patterns. The A. flavus/oryzae strains did not exhibit the high amylase activity detected in the commercial yellow koji strain (starter mold). However, their peptidase and lipase activities were generally higher than that of the koji isolates. We verified the safety of the traditionally fermented meju samples by analyzing the AGC breaking point pattern and the enzyme activities of A. flavus/oryzae strains isolated from the samples. The isolated strains could possibly be used as starter molds for soybean fermentation.

Aflatoxins are known to cause devastating acute and chronic effects in humans and animals. The objective of the study was to determine the influence of postharvest practices and storage conditions on aflatoxin contamination in maize in two counties. Aflatoxin levels in 144 maize samples from different maize storage conditions were determined. While sampling, a structured questionnaire was also administered to evaluate farmer’s postharvest practices. Makueni County had the highest percentage of aflatoxin positive samples with up to 174 ppb attributed to the long storage under unfavourable conditions. On the other hand, Baringo County had lower positivity associated with the harvesting season at the time of sample collection. The type of storage condition had a significant effect on the extent of contamination and accounted for 11% of the variation (R 2 = 0.11). Gunny bags were the most common type of storage condition and had the highest level of contamination in both the counties. Metallic bins had the lowest level of contamination. Aflatoxin G1 and G2 were predominant in samples from Baringo County, while aflatoxin B1 and B2 were predominant in samples from Makueni County. The study concluded that the type of storage condition significantly contributes to the aflatoxin contamination in the stored maize. Proper drying of maize to the recommended moisture content and subsequent storage in hermetic structures will reduce the cases of aflatoxin contamination.

This study was conducted in the mycotoxins laboratory, College of Agricultural Engineering Sciences, University of Baghdad, Iraq to evaluate the efficiency of silver nanoparticles manufactured locally and biologically by Aspergillus niger in preventing A. flavus to produce aflatoxin B1 (AFB1). The results of laboratory isolation showed that the companion of fungi genera were Aspergillus spp., Fusarium spp., Penicillum spp. and Rhizopus spp. at rates of 5.66, 14.91, 21.18 and 38.86% respectively. The highest frequency of A. flavus was 19.32%. The results of the TLC test showed that all isolates produced AFB1 in varying rates (%), and the Baghdad / Al-Youssifia isolate was the most productive of AFB1, since it has a largest spot area and most intense fluorescence under the chromatographic plate, hence given a symbolic name AFBY7. The results of HPLC showed that the toxin concentration in the AFBY7 isolate was 124.167 ppb. Also, the results showed the high efficiency of A. niger in the manufacture of silver nanoparticles, as the colour of solution changed from yellow to dark brown. On the other hand, the results of using locally and biologically manufactured silver nanoparticles in the storage experiment to prevent the fungus from producing toxin showed superiority of treatments (T) 0.4, 0.6 and 0.8 mg L-1, since AFB1 was 0.0 ppb compared to T0.2 (3.990 ppb). In addition, the results showed the efficiency of locally and biologically manufactured silver nanoparticles used in reducing AFB1 in the storage experiment of maize grains stored. So that, T0.6 and T0.8 were superior in reducing the AFB1 to 0.0, compared to T0.2 and T0.4, leading AFB1 to reach 10.230 and 5.180 ppb respectively.

Aspergillus flavus is an opportunistic pathogen of oilseed crops such as maize, peanut, cottonseed, and tree nuts and produces carcinogenic secondary metabolites known as aflatoxins during seed colonization. Aflatoxin contamination not only reduces the value of the produce but is also a health hazard to humans and animals. Previously, we observed inhibition of A. flavus aflatoxin biosynthesis on exposure to the marine bacterium Vibrio gazogenes (Vg). In this study, we used RNA sequencing to examine the transcriptional profiles of A. flavus treated with both live and heat-inactivated dead Vg and control samples. Fungal biomass, total accumulated aflatoxins, and expression profiles of genes constituting secondary metabolite biosynthetic gene clusters were determined at 24, 30, and 40 h after treatment. Statistically significant reductions in total aflatoxins were detected in Vg-treated samples as compared with control samples at 40 h. However, no statistical difference in fungal biomass was observed on these treatments. The Vg treatments were most effective on aflatoxin biosynthesis, as was reflected in significant downregulation of most genes in the aflatoxin gene cluster, including the aflatoxin pathway regulator gene, aflR. Along with aflatoxin genes, we also observed significant downregulation in some other secondary metabolite gene clusters, including cyclopiazonic acid and aflavarin, suggesting that the treatment could inhibit other secondary metabolites as well. Finally, a weighted gene correlation network analysis identified an upregulation of 10 genes that were most strongly associated with Vg-dependent aflatoxin inhibition and provide a novel starting point in understanding the mechanisms that result in this phenomenon. [Figure: see text] The author(s) have dedicated the work to the public domain under the Creative Commons CC0 “No Rights Reserved” license by waiving all of his or her rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law, 2022.

针对市场在售的调和油、玉米油、大豆油、花生油和菜籽油等食用植物油,随机购买20种共290份食用植物油样品。采用液相色谱法测定AFB1、AFB2、AFG1、AFG2以及OTA、OTB真菌毒素的含量，对食用植物油中黄曲霉毒素和赭曲霉毒素的污染水平和分布特征进行分析。结果表明：20种290份食用植物油样品中，有16种共67份样品存在不同程度的真菌毒素污染，总污染率达到23.1%，不同种类食用植物油污染呈现“多种类、共分布”的特点，其中AFG1污染率（14.8%）最高，其次为OTA（13.4%）。绝大多数阳性样本受1～4种真菌毒素污染，仅有少数阳性样本受真菌毒素污染数量达到5种。总体上食用植物油样品受到多种真菌毒素的混合污染情况比较严重，应引起一定的重视。 A total of 290 samples from 20 kinds of edible vegetable oils such as blended oil, corn oil, soybean oil, peanut oil, and rapeseed oil on the market were randomly purchased. Then, the contents of AFB1, AFB2, AFG1, AFG2, OTA and OTB were accurately determined by HPLC to investigate the pollution levels and distribution characteristics of aflatoxins and ochratoxins in edible vegetable oils. The results showed that among the 20 kinds of 290 edible vegetable oil samples, 16 kinds of 67 samples suffered from different degrees of mycotoxin contamination, and the total pollution rate reached 23.1%. The pollution of different kinds of edible vegetable oils presented the characteristics of multi-species and co-distribution. AFG1 had the highest pollution rate (14.8%), followed by OTA (13.4%). Besides, the vast majority of positive samples were contaminated by 1-4 kinds of mycotoxins, only a few positive samples were contaminated with 5 kinds of mycotoxins. Overall, the problem that the edible vegetable oil samples are polluted by multiple mycotoxins is serious and should be paid attention to.

In sub-Saharan Africa (SSA), millions of people depend on maize as a primary staple. However, maize consumers in SSA may be exposed to malnutrition due to vitamin A deficiency (VAD) and unsafe aflatoxin levels, which can lead to serious economic and public health problems. Provitamin A (PVA) biofortified maize has been developed to alleviate VAD and may have additional benefits such as reduced aflatoxin contamination. In this study, maize inbred testers with contrasting PVA content in grain were used to identify inbred lines with desirable combining ability for breeding to enhance their level of resistance to aflatoxin. Kernels of 120 PVA hybrids generated by crossing 60 PVA inbreds with varying levels of PVA (5.4 to 51.7 µg/g) and two testers (low and high PVA, 14.4 and 25.0 µg/g, respectively) were inoculated with a highly toxigenic strain of Aspergillus flavus. Aflatoxin had a negative genetic correlation with b-carotene (r = -0.29, p < 0.0001) and PVA (r = -0.23, p < 0.0001), indicating that hybrids with high PVA content accumulated less aflatoxin than those with low to medium PVA. Both general combining ability (GCA) and specific combining ability (SCA) effects of lines and testers were significant for aflatoxin accumulation, number of spores, PVA, and other carotenoids, with additive gene actions playing a prominent role in regulating the mode of inheritance (GCA/SCA ratio >0.5). Eight inbreds had combined significant negative GCA effects for aflatoxin accumulation and spore count with significant positive GCA effects for PVA. Five testcrosses had combined significant negative SCA effects for aflatoxin with significant positive SCA effects for PVA. The high PVA tester had significant negative GCA effects for aflatoxin, lutein, b-carotene, and PVA. The study identified lines that can be used as parents to develop superior hybrids with high PVA and reduced aflatoxin accumulation. Overall, the results point out the importance of testers in maize breeding programs to develop materials that can contribute to controlling aflatoxin contamination and reducing VAD. [ABSTRACT FROM AUTHOR]

The genus Fumaria is known for its strong medicinal effects. This research aimed to assess the phytochemical profile in various alkaloid fractions from Fumaria vaillantii Loisel and Fumaria asepala Boiss. grown in the Saveh region of the central province of Iran and their antifungal activity against Aspergillus flavus and Aspergillus niger was investigated in laboratory conditions. Also, the types and amount of alkaloid compounds and the effect of compounds on the expression of key genes of aflatoxin biosynthesis were conducted in order to investigate the ability of alkaloids of two Fumaria species to prepare environmentally friendly disinfectants. The total alkaloid contents were extracted using the soxhlet apparatus and then fractioned to perform GC-MS analysis. The effect of the extract of two species of Fumaria on the activity of Aspergillus flavus and Aspergillus niger were evaluated considering time factors (24 h, 48 h, and 72 h) and two concentrations (50 and 100 mg/mL). The total alkaloids of the leaf and fruit of F. vaillantii respectively contain (3.48 and 3.32 mgg-1DW,) and F. asupala (2.38 and 2.26 mgg-1DW, respectively). Fractions 1 and 4 of leaf and fruit showed the highest total alkaloid content, which was then mixed for GC-MS analysis; isoquinoline alkaloids, including protopine and sanguinarine; parfumine and allocryptopine accounted for the highest percentage of alkaloids in both species. F. vaillantii had a higher content of these alkaloids. The antifungal activity of fractions 1 and 4 reduced radial growth in both Aspergillus species. The alkaloid fractions at both species caused downregulation of aflatoxin B1 biosynthesis key genes (aflR, aflD, and aflM) in A. flavus and fumonisin B2, key genes (fum8 and fum6) in A. niger. The obtained data shows that alkaloid fractions extracted from Fumaria species, F. vaillantii, can be useful in the preparation of nature-friendly disinfectants for postharvest disinfection of fruits and vegetables. [ABSTRACT FROM AUTHOR]

The worldwide economic crop groundnut is infected by aflatoxigenic Aspergillus species (AAS), for which the soil is the main inoculum reservoir. Thus, to assess the spatiotemporal dynamics and association of AAS populations with biophysical factors, a total of 300 soil samples were collected (pre-planting, flowering, and physiological maturity) from five groundnut-growing districts in three zones of Ethiopia in 2019. AAS were isolated by dilution plating and their aflatoxin-producing potential determined using special media and UV light. Logistic regression models were used to analyze the effect of biophysical factors on AAS population dynamics. AAS populations varied across groundnut-growing districts and over time. In the Fedis district, the highest relative density of A. flavus non-sclerotia producer (NSP) strain (30.1%) and small sclerotia (S-strain) producers (26.1%) were recorded at pre-planting and maturity, whereas large sclerotia (L-strain) producers (4.1%) and A. parasiticus (11.6%) were recovered from the Kucha district at flowering and maturity, respectively. From Humbo and Kucha districts, the highest populations of A. flavus (10.3 × 104 cfu/g soil) and A. parasiticus (5.5 × 104 cfu/g soil), respectively, were recorded at maturity. The smallest population of AAS had a strong association with altitudes > 1640 m a.s.l., higher pH (> 7), low organic matter (0.572–1.378), and sand-sandy loam soil, factors which can be considered as management options. A. flavus S-strain prevalence had a positive relationship with increased soil temperature > 30 °C, which highlights the need for mitigation strategies as the climate warms. [ABSTRACT FROM AUTHOR]

Aflatoxins are immunosuppressive and carcinogenic secondary metabolites, produced by the filamentous ascomycete Aspergillus flavus, that are hazardous to animal and human health. In this study, we show that multiplexed host-induced gene silencing (HIGS) of Aspergillus flavus genes essential for fungal sporulation and aflatoxin production (nsdC, veA, aflR, and aflM) confers enhanced resistance to Aspergillus infection and aflatoxin contamination in groundnut (<20 ppb). Comparative proteomic analysis of contrasting groundnut genotypes (WT and near-isogenic HIGS lines) supported a better understanding of the molecular processes underlying the induced resistance and identified several groundnut metabolites that might play a significant role in resistance to Aspergillus infection and aflatoxin contamination. Fungal differentiation and pathogenicity proteins, including calmodulin, transcriptional activator-HacA, kynurenine 3-monooxygenase 2, VeA, VelC, and several aflatoxin pathway biosynthetic enzymes, were downregulated in Aspergillus infecting the HIGS lines. Additionally, in the resistant HIGS lines, a number of host resistance proteins associated with fatty acid metabolism were strongly induced, including phosphatidylinositol phosphate kinase, lysophosphatidic acyltransferase-5, palmitoyl-monogalactosyldiacylglycerol Δ-7 desaturase, ceramide kinase-related protein, sphingolipid Δ-8 desaturase, and phospholipase-D. Combined, this knowledge can be used for groundnut pre-breeding and breeding programs to provide a safe and secure food supply. [ABSTRACT FROM AUTHOR]

This study aimed to assess the prevalence, concentration of AFM1 in human breast milk, and to determine the risk of cancer for infants in sub-Saharan Africa. A systematic literature search was performed using PubMed, CINAHL, Web of science, global health, Cochrane, and Google Scholar electronic databases. A random-effects model was used to estimate the pooled prevalence and concentration of AFM1 in breast milk. The meta-analysis of 8 articles containing 9 studies showed the pooled prevalence of AFM1 in breast milk to be 56.18% (95% CI: 29.65–82.71) and the pooled concentration to be 31.12 ng/L (95% CI: 25.97–36.25). The cancer risk assessment indicated for both male and female 1-month infants in Sierra Leone (HI > 1) is high, and all the rest of the infants are free of risk (HI < 1). The pooled prevalence and mean concentration of AFM1 in breast milk is high. Monitoring of AFB1 concentration of commonly used foods will be of high value in reducing the burden of AFM1. [ABSTRACT FROM AUTHOR]

Aflatoxins (AFs) are mycotoxins produced by numerous species of Aspergillus. AFs contaminate agricultural commodities and thus feed and food including milk. So, this study aimed to assess the effect of the feeding type on the occurrence of AFM1 in camel milk. A total of 45 camel milk samples were obtained from the natural grazing herd in Shalatin (n= 20) and small scales farm breeding in Daraw (n= 25) in the period between September to December 2021. AFM1 levels were determined in these samples using the Vicam method. A significant difference was found between AFM1 levels in milk samples from herds of camels kept in a traditional environment (Natural grazing) and that in samples from camels in semi-intensive management systems (Onfarm). Of note, all was samples obtained from the nomadic area were free from AFM1, whereas, in the camel milk samples collected from a semi-intensive farm, AFM1 was reported in eight samples (32%) with 2 (8%) samples exceeding the EU Limits of 0.05 µg/ kg and 6 (24%) samples below such EU Limits. In conclusion, there was high contamination of milk samples obtained from camels reared on-farm with AFM1, while milk samples of camels naturally grazed were negative for AFM1. Therefore, milk hygiene from the farm, chilling, and distribution should be evaluated to reduce AFM1 levels in milk. [ABSTRACT FROM AUTHOR]

Meat products represent an important source of animal-derived protein. However, meat products are also implicated as a potential source of human exposure to some chemical residues such as aflatoxins (AFTs). The objectives of the present study were first to estimate the total AFTs content in the retailed meat products (beef luncheon, sausage, and pasterma) in Sharkia governorate, Egypt. Besides, dietary intake and potential health risks for the Egyptian population associated with the consumption of AFTs-contaminated meat products were further calculated. The obtained results revealed that pasterma had significantly the highest AFTs content, followed by luncheon, and sausage respectively. Calculation of the daily intakes of the total AFTs revealed that consumption of bovine meat products represents a potential source for dietary exposure to total AFTs. However, the calculation of the margin of exposure of total AFTs among the Egyptian population based on the average consumption of meat products revealed that such meat products might not possess a cancer risk among Egyptian adults and children. In general, excessive consumption of such meat products should be avoided for long-life protection against AFTs-related cancer risk. [ABSTRACT FROM AUTHOR]

One of the most desired and promising diets in Egypt is beef products. It is an enriched media for mycotoxins. The occurrence of aflatoxigenic Aspergillus species is inspected in commercial beef products, HPLC-quantitative amount of aflatoxin B1, B2, G1, and G2 excesses, and genetic identification of aflatoxin regulatory gene (aflR1) by conventional PCR. Two hundred and forty commercial products (minced meat, beef kofta, beef sausage, beef burger, beef luncheon, frozen meat, beef frozen liver, chicken luncheon, chicken burger, chicken frozen liver, mloha, and fesikh; n=20 for each) were collected from different markets at Aswan City, Egypt. Enumeration, isolation, and identification of mold species were carried to each sample. The amount of aflatoxins was measured using HPLC. Genetic identification of the aflR1 gene in Aspergillus was performed using PCR. Mloha samples recorded the highest total mold count whereas the beef luncheon recorded the lowest mould count. Four fungal genera were identified and Aspergillus spp recorded the main with an incidence of 25.8%. By PCR, the aflR1 gene was productively augmented in all the tested Aspergillus spp. The findings illustrated that among the samples that were examined; the prevalence of AFB1 was 65%, followed by AFG2 at 63%, AFB2 at 40%, and AFG1 at 30%. Additionally, mloha (724.2±14.6), poultry frozen liver (288±6.7), and beef frozen liver (91.6±12.2) had higher mean values of total aflatoxins contamination than other samples. Every sample that has been analyzed shows a positive correlation between the amount of reduced total mycotoxins found in the samples and the increased dose of gamma irradiation used to treat the samples. Conclusion: Aflatoxin is frequently linked to meat, poultry, and fish, as well as the products made from these foods. The production of aflatoxin in meat, as well as the products made from it, creates a danger to the public's health. Thus, the most effective way to prevent aflatoxigenic mould contamination during the product's production stages is to apply stringent hygienic standards when processing meat products and to use high-quality flavoring agents as spices. [ABSTRACT FROM AUTHOR]

The Aspergillus genus, the etiological agent of aspergillosis, is an important food contaminant and mycotoxin producer. Plant extracts and essential oils are a source of bioactive substances with antimicrobial potential that can be used instead of synthetic food preservatives. Species from the Lauraceae family and the Ocotea genus have been used as traditional medicinal herbs. Their essential oils can be nanoemulsified to enhance their stability and bioavailability and increase their use. Therefore, this study sought to prepare and characterize both nanoemulsion and essential oil from the Ocotea indecora's leaves, a native and endemic species from the Mata Atlântica forest in Brazil, and evaluate the activity against Aspergillus flavus RC 2054, Aspergillus parasiticus NRRL 2999, and Aspergillus westerdjikiae NRRL 3174. The products were added to Sabouraud Dextrose Agar at concentrations of 256, 512, 1024, 2048, and 4096 µg/mL. The strains were inoculated and incubated for up to 96 h with two daily measurements. The results did not show fungicidal activity under these conditions. A fungistatic effect, however, was observed. The nanoemulsion decreased the fungistatic concentration of the essential oil more than ten times, mainly in A. westerdjikiae. There were no significant changes in aflatoxin production. [ABSTRACT FROM AUTHOR]

Aflatoxins can easily grow and develop in many feed ingredients and influence the risk of several animal chronic diseases. The present study aimed to evaluate the effect of bentonite inclusion on performances, carcass yield, giblet, and blood constituents of broiler-fed contaminated aflatoxin through a metaanalytical approach. A database was developed based on scientific publications that were searched using several search tools such as Science Direct, PubMed, Scopus, and Google Scholar with "bentonite", "clay", "montmorillonite", "aflatoxin" and "broiler" as keywords. A total of 31 studies were retrieved and included in the analysis. Data analysis was based on the mixed model in which dietary bentonite inclusion was treated as the fixed effect and different studies were considered as random effects. Bentonite used were sodium bentonite, calcium bentonite, and montmorillonite form with levels ranging from 0 to 5%. Meanwhile, aflatoxin B1 levels in the diet ranged from 0 to 5 mg/kg. In this study, body weight gain, feed intake, and feed efficiency showed a linear increase (p<0.001) in all phases. The mortality rate linearly decreased (p<0.05) in the cumulative. Bentonite inclusion linearly increased (p<0.001) the carcass, breast, leg, bursa of Fabricius, and gizzard weight proportion. The percentage of kidney, liver, heart, pancreas, uric acid, and gamma-glutamyltransferase linearly decreased (p<0.001) by increasing bentonite levels. Dietary bentonite linearly increased (p<0.001) total protein, albumin, aspartate aminotransferase, calcium, glycogen, globulin, and hemoglobin (p<0.05). It was concluded that bentonites have the ability to eliminate the detrimental effects of aflatoxin, which may increase the broiler performance, carcass yield and prevent the change in abnormal giblet weight. For effectiveness and animal safety, the EFSA recommends using a maximum of 2% bentonite in the feed. [ABSTRACT FROM AUTHOR]

Aspergillus flavus infects stored grains and produces aflatoxins with highly toxic and carcinogenic properties, which are a threat to food safety and human health. Microbes and their bio-active compounds are considered effective agents to control A. flavus and aflatoxins during the pre- and post-harvest stages. Here, we reveal that Bacillus flexus strain TR-1 isolated from tea plant rhizosphere soil could efficiently inhibit the growth of A. flavus in dual culture tests through the production of antifungal volatiles. Additionally, these volatiles showed great inhibitory effects against A. flavus infection and aflatoxin production in maize and peanut with high water activity levels during storage. Scanning electron microscope analysis further proved that the conidia of A. flavus on the peanut surface treated with TR-1 volatiles could not germinate and were severely dehydrated and shrunken. Gas chromatography-tandem mass spectrometry revealed that five antifungal compounds were identified in the volatile profiles, including benzene, methyl thiolacetate, dimethyl disulfide, butanethioic acid S-methyl ester analogue, dimethyl trisulfide (DMTS), and S-methyl 3-methylbutanethioate. DMTS, with the best antagonistic effect, completely inhibited the growth of A. flavus at 10 μl−1 (compound volume/airspace volume). TR-1 also exhibited efficient and broad inhibitory effects against six other fungal pathogens. Overall, B. flexus strain TR-1 and its produced volatiles showed valid antifungal activity against phytopathogens. This study provides novel materials for the production of bioactive agents to control plant pathogens in stored grains. [ABSTRACT FROM AUTHOR]

Enteropathy is a pathophysiological condition characterized by decreased intestinal barrier function and absorption. Past studies have hypothesized that mycotoxins might impair children's growth by causing intestinal enteropathy, including interactions between mycotoxins and pathogens. We investigated the association of two mycotoxins, aflatoxin B1 (AFB1) and fumonisin B1 (FB1), independently and in conjunction with microbial pathogens, with fecal biomarkers of environmental enteropathy in children. As part of a larger MAL‐ED study, 196 children were recruited in Haydom, Tanzania, and followed for the first 36 months of life. The gut inflammation biomarkers myeloperoxidase (MPO), neopterin (NEO), and alpha‐1‐antitrypsin (A1AT) were analyzed in stool samples at 24 months; with mean concentrations 5332.5 ng/L MPO, 807.2 nmol/L NEO, and 0.18 mg/g A1AT. Forty‐eight children were measured for AFB1‐lys, with a mean of 5.30 (95% CI: 3.93‐6.66) pg/mg albumin; and 87 were measured for FB1, with a mean of 1.25 (95% CI: 0.72–1.76) ng/ml urine. Although the pathogens adenovirus and Campylobacter were associated with A1AT (p = 0.049) and NEO (p = 0.004), respectively, no association was observed between aflatoxin (MPO, p = 0.30; NEO, p = 0.08; A1AT, p = 0.24) or fumonisin (MPO, p = 0.38; NEO, p = 0.65; A1AT, p = 0.20) exposure and any gut inflammation biomarkers; nor were interactive effects found between mycotoxins and pathogens in contributing to intestinal enteropathy in this cohort. Although further studies are needed to confirm these results, it is possible that mycotoxins contribute to child growth impairment via mechanisms other than disrupting children's intestinal function. [ABSTRACT FROM AUTHOR]

Olive oil can be contaminated by fungal toxins; therefore, it is necessary to monitor the incidence of mycotoxins in this oil. In the present study, the pooled prevalence of detectable aflatoxin B1 (AFB1) in olive oil was evaluated using systematic review and meta-analysis approach from 1 January 1991 to 31 December 2020 (30 years study). The search was conducted via electronic databases involving Scopus, Web of Science, PubMed, Agris and Agricola. Synonyms were collected from combination of the MESH, Agrovoc and free text method. After screening and selection process of primary researches, full texts of eligible researches (46 studies) were evaluated and data of the nine studies as included researches were extracted. Random effect model was used to estimate the pooled prevalence of AFB1 in olive oil and weighing model of Dersimonian–Laired was applied. Summary measure of mycotoxin prevalence was estimated using Metaprop module of STATA and 95% confidence interval (CI) were calculated using the Binomial Exact Method. Pooled prevalence of AFB1 in olive oils were 32% (95% CI 8–56%) which means that 68% of olive oil were free of detectable contaminants of AFB1. Due to controversy over the results of primary studies, future researches and consequent subgroup analysis based on the main variables affecting the aflatoxins contamination in olive oil are recommended to achieve the conclusive results. [ABSTRACT FROM AUTHOR]

A comprehensive study of bacterial and biotoxin contaminants of ready-to-eat (RTE) foods in Nigeria is yet to be reported. Hence, this study applied 16S rRNA gene sequencing and a dilute-and-shoot LC-MS/MS method to profile bacteria and biotoxins, respectively, in 199 RTE food samples comprising eko (n = 30), bread (n = 30), shawarma (n = 35), aadun (n = 35), biscuits (n = 34), and kokoro (n = 35). A total of 631 bacterial isolates, clustered into seven operational taxonomic units, namely Acinetobacter, Bacillus, Klebsiella, Proteus and Kosakonia, Kurthia, and Yokenella, that are reported for the first time were recovered from the foods. One hundred and eleven metabolites comprising mycotoxins and other fungal metabolites, phytoestrogenic phenols, phytotoxins, and bacterial metabolites were detected in the foods. Aflatoxins, fumonisins, and ochratoxins contaminated only the artisanal foods (aadun, eko, and kokoro), while deoxynivalenol and zearalenone were found in industrially-processed foods (biscuit, bread, and shawarma), and citrinin was present in all foods except eko. Mean aflatoxin (39.0 µg/kg) in artisanal foods exceeded the 10 µg/kg regulatory limit adopted in Nigeria by threefold. Routine surveillance, especially at the informal markets; food hygiene and safety education to food processors and handlers; and sourcing of high-quality raw materials are proposed to enhance RTE food quality and safeguard consumer health. [ABSTRACT FROM AUTHOR]

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Aflatoxins (AFs) are fungal secondary metabolites frequently detected in soil that exhibit in vitro toxicity to certain soil microorganisms. However, microbial responses at different levels and in complex systems such as the soil environment have not been systematically studied. Therefore, we investigated multiple microbial responses in two different soils (sandy loam and clay) to aflatoxin B1 (AFB1) at environmentally relevant concentrations (0.5–500 µg kg−1) during a 28-day incubation. General microbial parameters for biomass (microbial biomass carbon and ergosterol), activity (glucose-induced and basal respiration), and catabolic functionality (substrate utilization patterns) were assessed. We observed minor and transient effects in both soils. In sandy loam, we found negative effects on activity and catabolic functionality with increased metabolic quotient, while clay soil exhibited stimulation for the same parameters, suggesting a hormetic effect due to reduced bioavailability through sorption onto clay minerals. Our results indicate that AFB1 does not pose a threat to general microbial indicators under the test conditions in soils without previous AF contamination. Given the toxic potential of AFs to specific microorganisms, further studies should investigate responses at higher taxonomic and functional levels in natural environments of aflatoxigenic fungi, such as tropical soils, and including additional physicochemical stressors. [ABSTRACT FROM AUTHOR]

Summary: The objective of this study was to quantify the carbon dioxide (CO2) produced by soybeans stored at 25 °C, with moisture contents (MC) of 12%, 14%, 18% and 22% (wet basis – w.b.), in a dynamic respiration measurement system, as well as to evaluate the accumulated dry matter loss (DML) throughout the stoichiometric relation between glucose and CO2, the safe storage time of stored soybeans and quantify the aflatoxins of the soybeans during storage. Also, the presence of aflatoxins B1, B2, G1 and G2 within soybeans were evaluated. Soybeans with high MC contributed to a significant increase in respiration rate (RR) and, consequently, greater DML during the storage. Compared to the MC of 12% (w.b.), the soybean RR increased 1.3, 2.8 and 19 times for soybeans at 14%, 18% and 22% (w.b.) respectively. Maximum storage time of soybean increased with the decrease of MC. Soybeans stored with 22% (w.b.) obtained high contamination by aflatoxin B1 and B2. [ABSTRACT FROM AUTHOR]

Certain viruses have been found to induce diverse biological pathways to carcinogenesis, evidenced by the presence of viral gene products in some tumors. Despite the fact that many fungal agents contain mycoviruses, until recently, their possible direct effects on human health, including carcinogenesis and leukemogenesis, had not been explored. In this regard, most studies of fungal agents have rightly concentrated on their mycotoxin formation and effects. Recently, the direct role of yeasts and fungi in the etiology of cancers, including leukemia, have been investigated. While greater attention has been placed on the carcinogenic effects of Candida, the role of filamentous fungi in carcinogenesis has also been explored. Recent findings from studies using the enzyme-linked immunosorbent assay (ELISA) technique indicate that the plasma of patients with acute lymphoblastic leukemia (ALL) uniformly contains antibodies for a certain mycovirus-containing Aspergillus flavus, while controls are negative. The exposure of mononuclear leukocytes from patients with ALL in full remission, and long-term survivors, to the product of this organism was reported to result in the re-development of typical genetics and cell surface phenotypes characteristic of active ALL. Mycoviruses are known to be able to significantly alter the biological characteristics and functions of their host. The possible carcinogenic and leukemogenic role of mycoviruses, with and without their host, needs to be further investigated. [ABSTRACT FROM AUTHOR]

Aflatoxin poisoning can harm broiler health and cause economic loss. Traditional diagnoses rely on human assessment, which is laborious and time-consuming. The objective of this study was to identify aflatoxin-poisoned broilers by analyzing three-dimensional accelerations collected by wearable accelerometers using five machine learning models, including K-Nearest Neighbor (KNN), Support Vector Classification (SVC), Decision Tree (DT), Random Forest (RF), and Gradient Boosting Decision Tree (GBDT). A total of 75 yellow feather broilers were reared in 3 pens (25 birds/pen) provided with 3 types of feed, including Normal (no Aflatoxin B 1 , control), AF50 (50 μg/kg Aflatoxin B 1 "AFB 1 "), and AF100 (100 μg/kg AFB 1). Lightweight tri-axial accelerometers were attached to 9 randomly selected birds (3 birds/pen) to continuously monitor the movement of broilers. Broiler behaviors were manually labelled as six categorises (sitting, standing, drinking, feeding, walking, and other) by observing the videos recorded by the camera installed in each pen. Different behavioral patterns were observed for AFB 1 poisoned birds as compared to that for sound birds. The poisoned broilers performed less standing, walking, feeding, and drinking behaviors, but more sitting behaviors. Five models could identify poisoned birds (AF100) with good accuracies (>86%). Higher misclassification rates were observed for all models in classifying AF50 and Normal groups. Among different models, the GBDT model performed best (97%) in differentiating Normal and AF50 broilers. In conclusion, three-dimensional accelerations and machine learning models could be used to identify aflatoxin-poisoned broilers with high accuracies. The findings could help in the development of viable systems that can identify diseased birds. • Poisoned broilers spent less time on feeding, drinking, walking, and standing. • Performances of models differ in identifying healthy and mild poisoning broilers. • The GBDT model performed better in identifying poisoned broilers. [ABSTRACT FROM AUTHOR]