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Applied Microbiology and Biotechnology May 2024Mycotoxin production by aflatoxin B1 (AFB1) -producing Aspergillus flavus Zt41 and sterigmatocystin (ST) -hyperproducer Aspergillus creber 2663 mold strains on corn and...
Mycotoxin production by aflatoxin B1 (AFB1) -producing Aspergillus flavus Zt41 and sterigmatocystin (ST) -hyperproducer Aspergillus creber 2663 mold strains on corn and rice starch, both of high purity and nearly identical amylose-amylopectin composition, as the only source of carbon, was studied. Scanning electron microscopy revealed average starch particle sizes of 4.54 ± 0.635 µm and 10.9 ± 2.78 µm, corresponding to surface area to volume ratios of 127 1/µm for rice starch and 0.49 1/µm for corn starch. Thus, a 2.5-fold difference in particle size correlated to a larger, 259-fold difference in surface area. To allow starch, a water-absorbing powder, to be used as a sole food source for Aspergillus strains, a special glass bead system was applied. AFB1 production of A. flavus Zt41 was determined to be 437.6 ± 128.4 ng/g and 90.0 ± 44.8 ng/g on rice and corn starch, respectively, while corresponding ST production levels by A. creber 2663 were 72.8 ± 10.0 µg/g and 26.8 ± 11.6 µg/g, indicating 3-fivefold higher mycotoxin levels on rice starch than on corn starch as sole carbon and energy sources. KEY POINTS: • A glass bead system ensuring the flow of air when studying powders was developed. • AFB1 and ST production of A. flavus and A. creber on rice and corn starches were studied. • 3-fivefold higher mycotoxin levels on rice starch than on corn starch were detected.
Topics: Oryza; Zea mays; Starch; Aspergillus; Aspergillus flavus; Aflatoxin B1; Sterigmatocystin; Microscopy, Electron, Scanning; Particle Size; Mycotoxins; Glass
PubMed: 38809353
DOI: 10.1007/s00253-024-13190-7 -
Frontiers in Veterinary Science 2024MicroRNAs (miRNAs) serve as key regulators in gene expression and play a crucial role in immune responses, holding a significant promise for diagnosing and managing... (Review)
Review
MicroRNAs (miRNAs) serve as key regulators in gene expression and play a crucial role in immune responses, holding a significant promise for diagnosing and managing diseases in farm animals. This review article summarizes current research on the role of miRNAs in various farm animal diseases and mycotoxicosis, highlighting their potential as biomarkers and using them for mitigation strategies. Through an extensive literature review, we focused on the impact of miRNAs in the pathogenesis of several farm animal diseases, including viral and bacterial infections and mycotoxicosis. They regulate gene expression by inducing mRNA deadenylation, decay, or translational inhibition, significantly impacting cellular processes and protein synthesis. The research revealed specific miRNAs associated with the diseases; for instance, gga-miR-M4 is crucial in Marek's disease, and gga-miR-375 tumor-suppressing function in Avian Leukosis. In swine disease such as Porcine Respiratory and Reproductive Syndrome (PRRS) and swine influenza, miRNAs like miR-155 and miR-21-3p emerged as key regulatory factors. Additionally, our review highlighted the interaction between miRNAs and mycotoxins, suggesting miRNAs can be used as a biomarker for mycotoxin exposure. For example, alterations in miRNA expression, such as the dysregulation observed in response to Aflatoxin B1 (AFB1) in chickens, may indicate potential mechanisms for toxin-induced changes in lipid metabolism leading to liver damage. Our findings highlight miRNAs potential for early disease detection and intervention in farm animal disease management, potentially reducing significant economic losses in agriculture. With only a fraction of miRNAs functionally characterized in farm animals, this review underlines more focused research on specific miRNAs altered in distinct diseases, using advanced technologies like CRISPR-Cas9 screening, single-cell sequencing, and integrated multi-omics approaches. Identifying specific miRNA targets offers a novel pathway for early disease detection and the development of mitigation strategies against mycotoxin exposure in farm animals.
PubMed: 38803799
DOI: 10.3389/fvets.2024.1372961 -
Sensors (Basel, Switzerland) May 2024Fluorescence induced by the excitation of a fluorophore with plane-polarized light has a different polarization depending on the size of the fluorophore-containing...
Fluorescence induced by the excitation of a fluorophore with plane-polarized light has a different polarization depending on the size of the fluorophore-containing reagent and the rate of its rotation. Based on this effect, many analytical systems have been implemented in which an analyte contained in a sample and labeled with a fluorophore (usually fluorescein) competes to bind to antibodies. Replacing antibodies in such assays with aptamers, low-cost and stable oligonucleotide receptors, is complicated because binding a fluorophore to them causes a less significant change in the polarization of emissions. This work proposes and characterizes the compounds of the reaction medium that improve analyte binding and reduce the mobility of the aptamer-fluorophore complex, providing a higher analytical signal and a lower detection limit. This study was conducted on aflatoxin B1 (AFB1), a ubiquitous toxicant contaminating foods of plant origins. Eight aptamers specific to AFB1 with the same binding site and different regions stabilizing their structures were compared for affinity, based on which the aptamer with 38 nucleotides in length was selected. The polymers that interact reversibly with oligonucleotides, such as poly-L-lysine and polyethylene glycol, were tested. It was found that they provide the desired reduction in the depolarization of emitted light as well as high concentrations of magnesium cations. In the selected optimal medium, AFB1 detection reached a limit of 1 ng/mL, which was 12 times lower than in the tris buffer commonly used for anti-AFB1 aptamers. The assay time was 30 min. This method is suitable for controlling almond samples according to the maximum permissible levels of their contamination by AFB1. The proposed approach could be applied to improve other aptamer-based analytical systems.
Topics: Aflatoxin B1; Aptamers, Nucleotide; Fluorescence Polarization; Polyelectrolytes; Biosensing Techniques; Polyamines; Limit of Detection; Fluorescent Dyes
PubMed: 38794084
DOI: 10.3390/s24103230 -
Materials (Basel, Switzerland) May 2024Clays are a class of porous materials; their surfaces are naturally covered by moisture. Weak thermal treatment may be considered practical to remove the water...
Clays are a class of porous materials; their surfaces are naturally covered by moisture. Weak thermal treatment may be considered practical to remove the water molecules, changing the surface properties and making the micro- and/or mesoporosities accessible to interact with other molecules. Herein, a modulated thermogravimetric analysis (MTGA) study of the moisture behavior on the structures of five, both fibrous and laminar, clay minerals is reported. The effect of the thermal treatment at 150 °C, which provokes the release of weakly adsorbed water molecules, was also investigated. The activation energies for the removal of the adsorbed water (Ea) were calculated, and they were found to be higher, namely, from 160 to 190 kJ mol, for fibrous clay minerals compared to lamellar structures, ranging in this latter case from 80 to 100 kJ mol. The thermal treatment enhances the rehydration in Na-montmorillonite, stevensite, and sepiolite structures with a decrease in the energy required to remove it, while Ea increases significantly in palygorskite (from 164 to 273 kJ mol). As a proof of concept, the MTGA results are statistically correlated, together with a full characterization of the physico-chemical properties of the five clay minerals, with the adsorption of two molecules, i.e., aflatoxin B1 (AFB1) and β-carotene. Herein, the amount of adsorbed molecules ranges from 12 to 97% for the former and from 22 to 35% for the latter, depending on the particular clay. The Ea was correlated with AFB1 adsorption with a Spearman score of -0.9. When the adsorbed water is forcibly removed, e.g., under vacuum conditions and high temperatures, the structure becomes the most important, decreasing the Spearman score between β-carotene and Ea to -0.6.
PubMed: 38793298
DOI: 10.3390/ma17102231 -
Molecules (Basel, Switzerland) May 2024Aflatoxins (AFs) including AFB, AFB, AFG and AFG are widely found in agriculture products, and AFB is considered one of the most toxic and harmful mycotoxins. Herein, a...
Aflatoxins (AFs) including AFB, AFB, AFG and AFG are widely found in agriculture products, and AFB is considered one of the most toxic and harmful mycotoxins. Herein, a highly sensitive (at the pg mL level) and group-specific enzyme-linked immunosorbent assay (ELISA) for the detection of AFB in agricultural and aquiculture products was developed. The AFB derivative containing a carboxylic group was synthesized and covalently linked to bovine serum albumin (BSA). The AFB-BSA conjugate was used as an immunogen to immunize mice. A high-quality monoclonal antibody (mAb) against AFB was produced by hybridoma technology, and the mAb-based ELISA for AFB was established. IC and limit of detection (LOD) of the ELISA for AFB were 90 pg mL and 18 pg mL, respectively. The cross-reactivities (CRs) of the assay with AFB, AFG, and AFG were 23.6%, 42.5%, and 1.9%, respectively, revealing some degree of group specificity. Corn flour, wheat flour, and crab roe samples spiked with different contents of AFB were subjected to ELISA procedures. The recoveries and relative standard deviation (RSD) of the ELISA for AFB in spiked samples were 78.3-116.6% and 1.49-13.21% ( = 3), respectively. Wheat flour samples spiked with the mixed AF (AFB, AFB, AFG, AFG) standard solution were measured by ELISA and LC-MS/MS simultaneously. It was demonstrated that the proposed ELISA can be used as a screening method for evaluation of AFs (AFB, AFB, AFG, AFG) in wheat flour samples.
Topics: Enzyme-Linked Immunosorbent Assay; Animals; Antibodies, Monoclonal; Aflatoxin B1; Mice; Food Contamination; Limit of Detection; Zea mays; Flour; Agriculture; Serum Albumin, Bovine
PubMed: 38792140
DOI: 10.3390/molecules29102280 -
International Journal of Molecular... May 2024The current review aims to outline and summarize the latest research on aflatoxin, with research studies describing natural, herbal and chemical compound applications in... (Review)
Review
AIMS
The current review aims to outline and summarize the latest research on aflatoxin, with research studies describing natural, herbal and chemical compound applications in animal (pig) models and in vitro cellular studies. Aflatoxin, a carcinogenic toxin metabolite, is produced by in humid environments, posing a threat to human health and crop production. The current treatment involves the prevention of exposure to aflatoxin and counteracting its harmful toxic effects, enabling survival and research studies on an antidote for aflatoxin.
OBJECTIVES
To summarize current research prospects and to outline the influence of aflatoxin on animal forage in farm production, food and crop processing. The research application of remedies to treat aflatoxin is undergoing development to pinpoint biochemical pathways responsible for aflatoxin effects transmission and actions of treatment.
SIGNIFICANCE
To underline the environmental stress of aflatoxin on meat and dairy products; to describe clinical syndromes associated with aflatoxicosis on human health that are counteracted with proposed treatment and preventive interventions. To understand how to improve the health of farm animals with feed conditions.
Topics: Animals; Animal Feed; Humans; Aflatoxin B1; Food Contamination; Aspergillus flavus
PubMed: 38791343
DOI: 10.3390/ijms25105305 -
Foods (Basel, Switzerland) May 2024This paper presents the first assessment of dietary exposure to aflatoxin M1 (AFM1) and associated health risks through milk and dairy product consumption in Armenia....
This paper presents the first assessment of dietary exposure to aflatoxin M1 (AFM1) and associated health risks through milk and dairy product consumption in Armenia. Data on AFM1 in raw milk were obtained from an annual residue monitoring program. Additionally, commonly consumed dairy products (pasteurized milk, cheese, sour cream, curd cheese) were sampled, considering the sources of raw milk used by dairy companies. Per capita consumption of raw milk was sourced from national food balance databases, while individual consumption data for dairy products was collected via a 24 h recall survey with 1400 adult respondents. Detectable levels of AFM1 were observed in 7.14% of raw milk samples (up to 0.334 μg/kg) and, albeit at lower amounts (up to 0.009 µg/kg), in 30% and 40% of sour cream and curd cheese, respectively. The AFM1 levels were lower than the national maximum permitted level (0.5 μg/kg); however, levels in raw milk exceeded the EU ML (0.05 μg/kg). The estimated margin of exposure values for dairy products indicated no significant risk, whereas a reasonable worst-case estimate, using the measurable levels of AFM1 in raw milk consumption indicated a potential public health concern. This study provides a scientific basis for evaluating aflatoxin issues in the Caucasus area.
PubMed: 38790817
DOI: 10.3390/foods13101518 -
Toxins May 2024Food-producing animals are exposed to mycotoxins through ingestion, inhalation, or dermal contact with contaminated materials. This exposure can lead to serious... (Review)
Review
Food-producing animals are exposed to mycotoxins through ingestion, inhalation, or dermal contact with contaminated materials. This exposure can lead to serious consequences for animal health, affects the cost and quality of livestock production, and can even impact human health through foods of animal origin. Therefore, controlling mycotoxin exposure in animals is of utmost importance. A systematic literature search was conducted in this study to retrieve the results of monitoring exposure to mycotoxins in food-producing animals over the last five years (2019-2023), considering both external exposure (analysis of feed) and internal exposure (analysis of biomarkers in biological matrices). The most commonly used analytical technique for both approaches is LC-MS/MS due to its capability for multidetection. Several mycotoxins, especially those that are regulated (ochratoxin A, zearalenone, deoxynivalenol, aflatoxins, fumonisins, T-2, and HT-2), along with some emerging mycotoxins (sterigmatocystin, nivalenol, beauvericin, enniantins among others), were studied in 13,818 feed samples worldwide and were typically detected at low levels, although they occasionally exceeded regulatory levels. The occurrence of multiple exposure is widespread. Regarding animal biomonitoring, the primary objective of the studies retrieved was to study mycotoxin metabolism after toxin administration. Some compounds have been suggested as biomarkers of exposure in the plasma, urine, and feces of animal species such as pigs and poultry. However, further research is required, including many other mycotoxins and animal species, such as cattle and sheep.
Topics: Animals; Mycotoxins; Animal Feed; Sheep; Food Contamination; Poultry; Swine; Cattle; Biological Monitoring; Livestock
PubMed: 38787070
DOI: 10.3390/toxins16050218 -
Toxins May 2024The fungal cell wall serves as the primary interface between fungi and their external environment, providing protection and facilitating interactions with the...
The fungal cell wall serves as the primary interface between fungi and their external environment, providing protection and facilitating interactions with the surroundings. Chitin is a vital structural element in fungal cell wall. Chitin deacetylase (CDA) can transform chitin into chitosan through deacetylation, providing various biological functions across fungal species. Although this modification is widespread in fungi, the biological functions of CDA enzymes in remain largely unexplored. In this study, we aimed to investigate the biofunctions of the CDA family in . The genome contains six annotated putative chitin deacetylases. We constructed knockout strains targeting each member of the CDA family, including Δ, Δ, Δ, Δ, Δ, and Δ. Functional analyses revealed that the deletion of CDA family members neither significantly affects the chitin content nor exhibits the expected chitin deacetylation function in . However, the Δ strain displayed distinct phenotypic characteristics compared to the wild-type (WT), including an increased conidia count, decreased mycelium production, heightened aflatoxin production, and impaired seed colonization. Subcellular localization experiments indicated the cellular localization of CDA6 protein within the cell wall of filaments. Moreover, our findings highlight the significance of the CBD1 and CBD2 structural domains in mediating the functional role of the CDA6 protein. Overall, we analyzed the gene functions of CDA family in , which contribute to a deeper understanding of the mechanisms underlying aflatoxin contamination and lay the groundwork for potential biocontrol strategies targeting .
Topics: Aspergillus flavus; Amidohydrolases; Aflatoxins; Fungal Proteins; Chitin; Cell Wall
PubMed: 38787069
DOI: 10.3390/toxins16050217 -
Toxins Apr 2024The aims of this study were (i) to determine the effect of an algoclay-based decontaminant on the oral availability of three mycotoxins (deoxynivalenol; DON, ochratoxin...
An Algoclay-Based Decontaminant Decreases Exposure to Aflatoxin B, Ochratoxin A, and Deoxynivalenol in a Toxicokinetic Model, as well as Supports Intestinal Morphology, and Decreases Liver Oxidative Stress in Broiler Chickens Fed a Diet Naturally Contaminated with Deoxynivalenol.
The aims of this study were (i) to determine the effect of an algoclay-based decontaminant on the oral availability of three mycotoxins (deoxynivalenol; DON, ochratoxin A; OTA, and aflatoxin B; AFB) using an oral bolus model and (ii) to determine the effect of this decontaminant on the performance, intestinal morphology, liver oxidative stress, and metabolism, in broiler chickens fed a diet naturally contaminated with DON. In experiment 1, sixteen 27-day-old male chickens (approximately 1.6 kg body weight; BW) were fasted for 12 h and then given a bolus containing either the mycotoxins (0.5 mg DON/kg BW, 0.25 mg OTA/kg BW, and 2.0 mg AFB/kg BW) alone ( = 8) or combined with the decontaminant (2.5 g decontaminant/kg feed; circa 240 mg/kg BW) ( = 8). Blood samples were taken between 0 h (before bolus administration) and 24 h post-administration for DON-3-sulphate, OTA, and AFB quantification in plasma. The algoclay decontaminant decreased the relative oral bioavailability of DON (39.9%), OTA (44.3%), and AFB (64.1%). In experiment 2, one-day-old male Ross broilers ( = 600) were divided into three treatments with ten replicates. Each replicate was a pen with 20 birds. The broiler chickens were fed a control diet with negligible levels of DON (0.19-0.25 mg/kg) or diets naturally contaminated with moderate levels of DON (2.60-2.91 mg/kg), either supplemented or not with an algoclay-based decontaminant (2 g/kg diet). Jejunum villus damage was observed on day 28, followed by villus shortening on d37 in broiler chickens fed the DON-contaminated diet. This negative effect was not observed when the DON-contaminated diet was supplemented with the algoclay-based decontaminant. On d37, the mRNA expression of glutathione synthetase was significantly increased in the liver of broiler chickens fed the DON-contaminated diet. However, its expression was similar to the control when the birds were fed the DON-contaminated diet supplemented with the algoclay-based decontaminant. In conclusion, the algoclay-based decontaminant reduced the systemic exposure of broiler chickens to DON, OTA, and AFB in a single oral bolus model. This can be attributed to the binding of the mycotoxins in the gastrointestinal tract. Moreover, dietary contamination with DON at levels between 2.69 and 2.91 mg/kg did not impair production performance but had a negative impact on broiler chicken intestinal morphology and the liver redox system. When the algoclay-based decontaminant was added to the diet, the harm caused by DON was no longer observed. This correlates with the results obtained in the toxicokinetic assay and can be attributed to a decreased absorption of DON.
Topics: Animals; Chickens; Trichothecenes; Oxidative Stress; Male; Ochratoxins; Liver; Aflatoxin B1; Animal Feed; Food Contamination; Intestines; Toxicokinetics; Diet; Aluminum Silicates
PubMed: 38787059
DOI: 10.3390/toxins16050207