Ochratoxin Research Articles

Deciphering the antiochratoxigenic activity of plant extracts and Debaryomyces hansenii against Penicillium nordicum in a “chorizo”-based medium by proteomic analysis

Penicillium nordicum is the main ochratoxin A (OTA)-producing species on the surface of dry-fermented sausages, such as the “chorizo”. New antifungal strategies are being developed using biocontrol agents (BCAs), such as plant extracts and native microorganisms. This work aimed to evaluate the antiochratoxigenic capacity and the causative modes of action of BCAs (rosemary essential oil (REO), acorn shell extract and the yeast Debaryomyces hansenii (Dh)) in a “chorizo”-based medium (Ch-DS). BCAs were inoculated on Ch-DS together with P. nordicum and incubated at 12 °C for 15 days to collect mycelia for OTA analyses and comparative proteomics. Both REO and Dh alone decreased OTA accumulation up to 99% and affected the abundance of P. nordicum proteins linked to cell wall organisation, synthesis of OTA-related metabolites and ergosterol synthesis. It is worth highlighting the increased abundance of an amidase by REO, matching with the decrease in OTA. The use of REO and Dh as BCAs could be an effective strategy to reduce the OTA hazard in the meat industry. Based on their not fully coincident modes of action, their combined application could be of interest in “chorizo” to maximise their potential against ochratoxigenic strains.

Assembly of stabilized dendritic magnetic ferric-silver nanocomposite with gold nanoparticles for sensitive detection of ochratoxin A using SERS aptasensor

Ochratoxin A (OTA) contamination poses a significant global threat to food security, necessitating the development of rapid and effective detection methods. A dendritic magnetic silver nanocomposite (Fe3O4@SiO2@Ag) tagged with aptamers acted as surface-enhanced Raman scattering (SERS) substrates and capture probes. Further, gold nanoparticles (AuNPs) were functionalized with cDNA and utilized as Raman-enhanced probes (cDNA-AuNPs). The SERS aptasensor which was constructed by the assembly of Apt-Fe3O4@SiO2@Ag and cDNA-AuNPs attained the accuracy quantitative and extremely sensitive detection of OTA. An inverse relationship between the concentration of OTA standard solution and Raman intensity of the characteristic peak at 1080 cm−1 was established in the range of 0.01–5 ng/mL, with an LOD of 0.0074 ng/mL. Furthermore, for positive corn samples, the coincidence rate between the established SERS aptamer sensor and HPLC was in the range of 92.67–121.10%, demonstrating tremendous promise for future practical application through other crops.

A highly efficient homogeneous electrochemiluminescence aptasensor for amplified analysis of Ochratoxin A

Ochratoxin A (OTA) is a toxic thermostable subordinate metabolite produced by Aspergillus and Penicillium species. OTA contamination occurs in a wide range of foods and poses a threat to human health. Therefore, it is crucial to construct a sensitive and reliable method for analyzing OTA to ensure food safety. Herein, a simple yet sensitive ECL aptasensor has been constructed for OTA detection through the synergistic amplification between catalytic DNA assembly (CDA) and exonuclease III (Exo III)-initiated recycling reaction. The anti-OTA aptamer-encoded sensing unit specifically recognizes target OTA and leads to the release of the trigger sequence for stimulating the CDA circuit, yielding numerous dsDNA hybrids that carry the re-assembled S strand. Each of the as-assembled S sequence could hybridize with the Ru(bpy)32+-labeled substrate (S*-Ru) to produce a T-shape structure with a blunt 3′ terminus, which can be hydrolyzed and digested by Exo III, leading to the liberation of mononucleotide functionalized with Ru(bpy)32+ (Ru). Simultaneously, the S sequence was released and hybridized with another S*-Ru sequence for initiating the subsequent cycle of Exo III-stimulated digestion process until all the S*-Ru strands are digested, thus resulting in the generation of numerous Ru and remarkably amplified ECL signals for high-performance detection of OTA. With reliable analyte recognition and high signal gain, the ECL system enabled the high-performance and accurate determination of OTA in buffer, wheat, and wine samples, thus showing considerable potential applications for highly efficient determination of contaminants of low concentration in foodstuff.

Integrating CdIn2S4 semiconductors with S-vacancy MoS2 nanosheets to fabricate a multi-channel aptasensing chip for photoelectrochemical detection of multiple mycotoxins

BackgroundThe importance of multi-target simultaneous detection lies in its ability to significantly boost detection efficiency, making it invaluable for rapid and cost-effective testing. Photoelectrochemical (PEC) sensors have emerged as promising candidates for detecting harmful substances and biomarkers, attributable to their unparalleled sensitivity, minimal background signal, cost-effectiveness, equipment simplicity, and outstanding repeatability. However, designing an effective multi-target detection strategy remains a challenging task in the PEC sensing field. Consequently, there is a pressing need to address the development of PEC sensors capable of simultaneously detecting multiple targets. ResultsCdIn2S4/V–MoS2 heterojunctions were successfully prepared via a hydrothermal method. These heterojunctions exhibited a high photocurrent intensity, representing a 1.53-fold enhancement compared to CdIn2S4 alone. Next, we designed a multi-channel aptasensing chip using ITO as the substrate. Three working electrodes were created via laser etching and subsequently modified with CdIn2S4/V–MoS2 heterojunctions. Thiolated aptamers were then self-assembled onto the CdIn2S4/V–MoS2 heterojunctions via covalent bonds, serving as recognition tool. By empolying the CdIn2S4/V–MoS2 heterojunctions as the sensing platform and aptamers as recognition tool, we successfully developed a disposable aptasensing chip for the simultaneous PEC detection of three typical mycotoxins (aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN)). This aptasensing chip exhibited wide detection range for AFB1 (0.05–50 ng/mL), OTA (0.05–500 ng/mL), and ZEN (0.1–250 ng/mL). Furthermore, it demonstrated ultra-low detection limits of 0.017 ng/mL for AFB1, 0.016 ng/mL for OTA, and 0.033 ng/mL for ZEN. Significance and noveltyThe aptasensing chip stands out for its cost-effectiveness, simplicity of fabrication, and multi-channel capabilities. The versatility and practicality enable it to serve as a powerful platform for designing multi-channel PEC aptasensors. With its ability to detect multiple targets with high sensitivity and specificity, the aptasensing chip holds immense potential for applications across diverse fields, such as environmental monitoring, clinical diagnostics, and food safety monitoring, where multi-target detection is crucial.

Membrane Receptor-Mediated Disruption of Cellular Homeostasis: Changes in Intracellular Signaling Pathways Increase the Toxicity of Ochratoxin A.

Organisms maintain their cellular homeostatic balance by interacting with their environment through the use of their cell surface receptors. Membrane based receptors such as the transforming growth factor β receptor (TGFR), the prolactin receptor (PRLR), and hepatocyte growth factor receptor (HGFR), along with their associated signaling cascade, play significant roles in retaining cellular homeostasis. While these receptors and related signaling pathways are essential for health of cell and organism, their dysregulation can lead to imbalance in cell function with severe pathological conditions such as cell death or cancer. Ochratoxin A (OTA) can disrupt cellular homeostasis by altering expression levels of these receptors and/or receptor-associated intracellular downstream signaling modulators and/or pattern and levels of their phosphorylation/dephosphorylation. Recent studies have shown that the activity of the TGFR, the PRLR, and HGFR and their associated signaling cascades change upon OTA exposure. A critical evaluation of these findings suggests that while increased activity of the HGFR and TGFR signaling pathways leads to an increase in cell survival and fibrosis, decreased activity of the PRLR signaling pathway leads to tissue damage. This review explores the roles of these receptors in OTA-related pathologies and effects on cellular homeostasis.

Embryo injected with Ochratoxin A induced jejunum injury in ducklings by activating the TLR4 signaling pathway: Involvement of intestinal microbiota

Ochratoxin A (OTA) is a common mycotoxin that causes intestinal injury in humans and various animal species. OTA may lead to intestinal injury in offspring due to the maternal effect. The aim of this study was to investigate the mechanism of embryo injected with OTA induced jejunum injury in ducklings. The results showed that OTA disrupted the jejunum tight junctions in hatching ducklings, and promoted the secretion of inflammatory cytokines. And this inflammatory response was caused by the activation of the TLR4 signaling pathway. Moreover, embryo injected with OTA could cause damage to the intestinal barrier in 21-day-old ducks, characterized by shortened villi, crypt hyperplasia, disrupted intestinal tight junctions, increased level of LPS in the jejunum, activation of the TLR4 signaling pathway, and increased levels of pro-inflammatory cytokines. Meanwhile, OTA induced oxidative stress in the jejunum. And dysbiosis of gut microbiota was mainly characterized by an increased the relative abundance of Bacteroides, Megamonas, Fournierella, and decreased the relative abundance of Alistipes and Weissella. Interestingly, embryo injected with OTA did not induce these changes in the jejunum of antibiotics-treated 21-day-old ducks. In conclusion, embryo injected with OTA induced jejunum injury in ducklings by activating the TLR4 signaling pathway, which involvement of intestinal microbiota.

Involvement of multiple epigenetic mechanisms by altered DNA methylation from the early stage of renal carcinogenesis before proliferative lesion formation upon repeated administration of ochratoxin A

Ochratoxin A (OTA) is a rat renal carcinogen that induces karyomegaly and micronuclei in proximal tubular epithelial cells (PTECs). We previously performed comprehensive gene profiling of alterations in promoter-region methylation and gene expression in PTECs of rats treated with OTA for 13 weeks. The OTA-specific gene profile was obtained by excluding genes showing expression changes similar to those upon treatment with 3-chloro-1,2-propanediol, a renal carcinogen not inducing karyomegaly. In this study, we validated the candidate genes using methylated DNA enrichment PCR and real-time RT-PCR, and identified Gen1, Anxa3, Cdkn1a, and Osm as genes showing OTA-specific epigenetic changes. These genes and related molecules were subjected to gene expression and immunohistochemical analyses in the PTECs of rats treated with OTA, other renal carcinogens, or non-carcinogenic renal toxicants for 4 or 13 weeks. Cdkn1a upregulation and increase of p21WAF1/CIP1+ karyomegalic PTECs were observed with OTA, matching the findings associated with micronucleus-inducing carcinogens. This suggested that the increase of p21WAF1/CIP1+ karyomegalic PTECs is linked to micronucleus formation, which in turn accelerates chromosomal instability. The upregulation of Cdkn1a-related genes with OTA suggests the acquisition of a senescence-associated secretory phenotype, which promotes the establishment of a carcinogenic environment. Meanwhile, OTA specifically caused a decrease of GEN1+ PTECs reflecting Gen1 downregulation and an increase of ANXA3+ PTECs reflecting Anxa3 upregulation, as well as Osm upregulation. OTA may efficiently disrupt pathways for repairing the DNA double-strand breaks that it itself causes, via Gen1 downregulation, and enhance cell proliferation through the upregulation of Anxa3 and Osm. This may exacerbate the chromosomal instability from the early stage of OTA-induced renal carcinogenesis before proliferative lesions form. OTA may cause renal carcinogenesis involving multiple epigenetic mechanisms.

Surface tailoring of Graphene via silicon co-doping with Group 15 Elements for the Detection of Ochratoxin (OTX): An Insilco Investigation

Today, the world is finding it challenging to recover from microorganism pollution, which is a concern for our world's safety and people's well-being. Ochratoxin is a well-known mycotoxin caused by Aspergillus and Penicillium species, and the contamination of crops, soil, air, and water by this toxin has far-reaching consequences that affect ecosystems and potentially lead to chronic health issues in humans and animals. However, the environmental impact of ochratoxins (OTXs) today is a multiple challenge that extends across agricultural landscapes and ecosystems. Herein, we intend to resolve these concerns through sensor technology, whereby this toxin can be adsorbed from various environments. We re-engineered graphene nanomaterials with codoped silicon and group 15 elements as sensors for OTX through a computational study. The modification of graphene with the mentioned elements demonstrates strong stability characteristics. This is evidenced by the transitions from π antibonding to another π antibonding, lone pair (LP) to LP, and π antibonding via the NBO study. Silicon-doped graphene showed good adsorption of OTX, and the adsorption strength further increased after the 15-group elements were doped with Si@GP, providing a more suitable option. The adsorption strength increased in the following order: OTX-P-Si@GP (-2.3113 eV), OTX-As-Si@GP (-3.2595 eV), OTX-Sb-Si@GP (-5.4025 eV), and OTX-Bi-Si@GP (-6.6488 eV). Therefore, the modification of Si@GP with 15 group elements is unique for ochratoxin adsorption such that as the electronegativity of 15 group elements decreases, the surface becomes more suitable for the adsorption of OTX. Interestingly, the results of this research suggested that the studied surfaces can be a potential candidate in the application of OTX sensors, thus this study can be beneficial to industries and experimental researchers.

Comparative Assessment of Thermal Treatment for Mycotoxins Adsorption Potential of Indigenous Clay from Balochistan, Pakistan

The current study aimed to investigate the physical characteristics and mycotoxins (Aflatoxins B1 (AFB1) and Ochratoxin A (OTA) binding efficacy of indigenous clays before and after thermal treatment (calcination). The clay sample was collected from various mountainous areas of Balochistan, Pakistan. The samples were ground into a 360 mesh size and thermally treated initially at 200 °C and thereafter at 800 °C for 30 minutes with the help of a muffle furnace. XRD analysis revealed montmorillonite as the most commonly occurring group of clay minerals. The thermally treated clays showed that the binding potential was significantly (P <0.05) improved against mycotoxins. Enzyme-linked immunoassay (ELISA) determination showed significantly (P <0.05) lower residual concentrations of mycotoxins in thermally treated clays. The mycotoxin adsorption capacity (%) was found to be higher in the clay samples procured from Pishin (79.17% for AFB1 and 67.34% for OTA) and Bolan (62.0% for AFB1 and 50.17% for OTA). Calcination also caused a significant decrease in Cation Exchange Capacity (CEC) in some clay samples. Overall, the findings of the study showed that thermal treatment enhanced the in vitro binding capacity of clays against the studied mycotoxins. An increase in the mycotoxin binding capacity of the clays not only reduces the bioavailability of mycotoxins but also decreases their toxic effects. It is recommended that Balochistan clay have the potential to be used as an adsorbent for the removal of mycotoxins from the environment and animal feeds.

A Portable Automated Microfluidic Platform for Point-of-Care Testing for Multiple Mycotoxins in Wine.

Food safety requires point-of-care testing (POCT) for mycotoxins, since their presence in wine significantly impacts the wine industry and poses a severe threat to human life. Traditional detection methods are usually limited to detecting one mycotoxin and cannot achieve high-throughput, automated, and rapid quantitative analysis of multiple mycotoxins in real samples. Here, we propose a portable automated microfluidic platform (PAMP) integrating a chemiluminescence (CL) imaging system and a microfluidic chip to realize POCT for multiple mycotoxins in real samples, simplifying complex manual operations, shortening the detection time, and improving the detection sensitivity. Specially, silicone films were used as substrates on microfluidic chips to incubate mycotoxin conjugations, and the streptavidin-biotin (SA-B) system and an indirect immunoassay were implemented on silicone films to improve the sensitivity of reaction results. Interestingly, these methods significantly improved detection results, resulting in sensitive detection of mycotoxins, including zearalenone (ZEA) ranging from 1 to 32 ng/mL, aflatoxin B1 (AFB1) ranging from 0.2 to 6.4 ng/mL, and ochratoxin A (OTA) ranging from 2 to 64 ng/mL. The recovery of samples reached 91.39-109.14%, which verified the reliability and practicability of the PAMP. This PAMP enables sensitive and rapid detection of multiple mycotoxins in markets or wineries that lack advanced laboratory facilities. Therefore, it is essential to develop a portable microfluidic platform for POCT to detect mycotoxins in real samples.

Phytic acid alleviates ochratoxin A-induced renal damage in chicks by modulating ferroptosis and the structure of the intestinal microbiota

Phytic acid (PA) is a natural antioxidant with various biological activities, providing protective effects in multiple animals. Ochratoxin A (OTA) is a mold toxin commonly found in feed, which induces multi-organ damage, with kidney being the target organ of its toxicity. This study investigates the protective effects of PA on OTA-induced renal damage and its potential mechanisms in chicks. The results demonstrates that PA treatment restores OTA-induced renal pathological injuries, reverses the diminished activities of antioxidant enzymes, reduces the accumulation of malondialdehyde, and normalizes the expression of pro-inflammatory cytokines, which confirms that PA can alleviate OTA-induced renal damage. Further investigations reveal that OTA-induced renal injury accompanied by an increase in tissue iron content and the transcription levels of ferroptosis-related genes (TFR, ACSL4, and HO-1), and a decrease in the levels of SLC7A11 and GPX4. PA treatment reverses all these effects, indicating that PA mitigates OTA-induced renal ferroptosis. Moreover, PA supplementation improves intestinal morphology and mucosal function, corrects OTA-induced changes in the intestinal microbiota. Besides, PA microbiota transplantation alleviates renal inflammation and oxidative stress caused by OTA. In conclusion, PA plays a protective role against renal damage through the regulation of ferroptosis and the intestinal microbiota, possibly providing novel insights into the control and prevention of OTA-related nephrotoxicity.

Self-designed portable dual-mode fluorescence device with custom python-based analysis software for rapid detection via dual-color FRET aptasensor with IoT capabilities

An innovative aptasensor incorporating MoS2-modified bicolor quantum dots and a portable spectrometer, designed for the simultaneous detection of ochratoxin A (OTA) and aflatoxin B1 (AFB1) in corn was developed. Carbon dots and CdZnTe quantum dots were as nano-donors to label OTA and AFB1 aptamers, respectively. These labeled aptamers were subsequently attached to MoS2 receptors, enabling fluorescence resonance energy transfer (FRET). With targets, the labeled aptamers detached from the nano-donors, thereby disrupting the FRET process and resulting in fluorescence recovery. Furthermore, a portable dual-mode fluorescence detection system, complemented with customized python-based analysis software, was developed to facilitate rapid and convenient detection using this dual-color FRET aptasensor. The developed host program is connected to the spectrometer and transmits data to the cloud, enabling the device to have Internet of Things (IoT) characteristics. Connected to the cloud, this IoT-enabled device offers convenient and reliable fungal toxin detection for food safety.

Simultaneous detection of multiple mycotoxins using MXene-based electrochemical aptasensor array and a self-developed multi-channel portable device

In response to the pressing need for highly efficient simultaneous detection of multiple mycotoxins, which are often found co-occurring in food raw materials and feed, an MXene-based electrochemical aptasensor array (MBEAA) was developed. This aptasensor array utilizes high-specificity aptamers as recognition elements, enabling the capture of electrical signal changes in the presence of target mycotoxins. Based on this platform, a multi-channel portable electrochemical device, enabling rapid, cost-effective, and simultaneous detection of aflatoxin B1 (AFB1), ochratoxin A (OTA), and zealenone (ZEN) was further developed. The developed system boasts a wide detection range of 1.0 × 10−1 to 10.0 ng mL−1, with remarkable performance characterized by ultra-low detection limits of 41.2 pg mL−1, 27.6 pg mL−1, and 33.0 pg mL−1 for AFB1, OTA, and ZEN, respectively. Successfully applied in corn samples, this method offers a portable, easy-to-operate, and cost-effective solution for simultaneous multi-mycotoxin detection. Moreover, the application of the self-developed detection system could be expanded for simultaneous detection of many different targets when their specific aptamers or antibodies were available.

Central role of Sigma-1 receptor in ochratoxin A-induced ferroptosis.

Ochratoxin A (OTA), a secondary fungal metabolite known for its nephrotoxic effects, is prevalent in various feeds and food items. Our recent study suggests that OTA-induced nephrotoxicity is linked to the Sigma-1 receptor (Sig-1R)-mediated mitochondrial pathway apoptosis in human proximal tubule epithelial-originated kidney-2 (HK-2) cells. However, the contribution of Sig-1R to OTA-induced nephrotoxicity involving other forms of regulated cell death, such as ferroptosis, remains unexplored. In this investigation, cell viability, malondialdehyde (MDA) levels, glutathione (GSH) levels, and protein expressions in HK-2 cells treated with OTA and/or Ferrostatin-1/blarcamesine hydrochloride/BD1063 dihydrochloride were assessed. The results indicate that a 24h-treatment with 1μM OTA significantly induces ferroptosis by inhibiting Sig-1R, subsequently promoting nuclear receptor coactivator 4 (NCOA4), long-chain fatty acid-CoA ligase 4 (ACSL4), arachidonate 5-lipoxygenase (ALOX5), autophagy protein 5 (ATG5), and ATG7, inhibiting ferritin heavy chain (FTH1), solute carrier family 7 member 11 (SLC7A11/xCT), glutathione peroxidase 4 (GPX4), peroxiredoxin 6 (PRDX6), and ferroptosis suppressor protein 1 (FSP1), reducing GSH levels, and increasing MDA levels (P < 0.05). In conclusion, OTA induces ferroptosis by inhibiting Sig-1R, subsequently promoting ferritinophagy, inhibiting GPX4/FSP1 antioxidant systems, reducing GSH levels, and ultimately increasing lipid peroxidation levels in vitro.

Assessment of Mycotoxin Exposure and Associated Risk in Pregnant Dutch Women: The Human Biomonitoring Approach.

Mycotoxins are toxic secondary metabolites produced by various fungi that can contaminate food crops, which, in turn, may lead to human exposure. Chronic exposure to mycotoxins can cause adverse health effects including reproductive and developmental toxicity. Pregnant women and their foetuses present a vulnerable group for exposure to mycotoxins that can cross the placenta. Human biomonitoring of mycotoxins provides a real-life approach to estimate internal exposure. In this pilot study, 24-h urine samples from 36 pregnant Dutch women were analysed for aflatoxin M1 (AFM1), total deoxynivalenol (DON), de-epoxy-deoxynivalenol (DOM-1), total zearalenone (ZEN), total α-zearalenol (α-ZEL), total β-zearalenol (β-ZEL) and total zearalanone (ZAN), where 'total' refers to mycotoxins and their conjugated forms. Serum samples from these women were analysed for fumonisin B1 (FB1) and ochratoxin A (OTA). All samples were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The most prevalent mycotoxins were total DON, total ZEN and OTA, with a detection frequency of 100%. DOM-1, total α-ZEL and total β-ZEL were detected but to a lesser extent, while AFM1, total ZAN and FB1 were undetected. Median concentrations were 4.75 μg total DON/L, 0.0350 μg DOM-1/L, 0.0413 μg total ZEN/L, 0.0379 μg total α-ZEL/L, 0.0189 μg total β-ZEL/L, and 0.121 μg OTA/L. The calculated median concentration for total ZEN and its metabolites was 0.105 μg/L. Based on two separate risk assessment approaches, total DON exposure in this group was considered to be of low concern. Similarly, exposure to total ZEN and its metabolites in this group was of low concern. For OTA, the risk of non-neoplastic effects was of low concern based on exposure in this group, and the risk of neoplastic effects was of low concern in the majority of participants in this group. The findings of this pilot study confirm the presence of mycotoxins in the urine and serum of pregnant Dutch women, with total DON, total ZEN, and OTA most frequently detected. Exposure to all measured mycotoxins was considered to be of low concern in this group, except for exposure to OTA, which was of low concern for the majority of participants. The study's findings offer valuable insights but should be confirmed using a larger and more diverse sample of the Dutch general population.

The toxigenic potential of phytopathogenic fungi isolated from pineapple Ananas comosus (L.) Merr. var MD-2 associated with postharvest rot disease

MD-2 pineapple fruits are susceptible to phytopathogenic fungi producing mycotoxins, which can cause up to 40% of postharvest losses. This study aimed to isolate and identify fungi associated with postharvest diseases on pineapple fruits, and mycotoxins produced. Thirty-seven fungal strains were isolated from the pineapple fruit at the full maturation stage. About 51.37% of the fungal strains identified were belonging to Aspergillus. Those fungi that caused diseases in pineapple fruits were Aspergillus sclerotioniger, A. tubingensis, Penicillium citrinum, Fusarium equiseti, Trichoderma asperelliodes and T. asperellum. Aspergillus niger produced the highest ochratoxin A (OTA), scoring 85.51 µg/kg. On the other hand, P. citrinum scored 94.06 µg/kg, the highest quantity of total aflatoxins. This is the first report demonstrating mycotoxigenic fungi’s potential to cause infections in postharvest pineapple fruits in Mexico.

Rational design of molecularly imprinted electrochemical sensor based on Nb2C-MWCNTs heterostructures for highly sensitive and selective detection of Ochratoxin a

Developing an efficient method for screening Ochratoxin A (OTA) in agriculture products is vital to ensure food safety and human health. However, the complex food matrix seriously affects the sensitivity and accuracy. To address this issue, we designed a novel molecularly imprinted polymer (MIP) electrochemical sensor based on multiwalled carbon nanotube-modified niobium carbide (Nb2C-MWCNTs) with the aid of the density functional theory (DFT). In this design, a glassy carbon electrode (GCE) was first modified by Nb2C-MWCNTs heterostructure. Afterward, the MIP layer was prepared, with ortho-toluidine as a functional monomer selected via DFT and OTA acting as a template on the surface of Nb2C-MWCNTs/GCE using in-situ electropolymerization. Electrochemical tests and physical characterization revealed that Nb2C-MWCNTs improved the sensor's active surface area and electron transmission capacity. Nb2C-MWCNTs had a good synergistic effect on MIP, endowing the sensor with high sensitivity and specific recognition of OTA in complex food matrix systems. The MIP sensor showed a wide linear range from 0.04 to 10.0 μM with a limit of detection (LOD) of 3.6 nM. Moreover, it presented good repeatability and stability for its highly antifouling effect on OTA. In real sample analysis, the recoveries, ranging from 89.77% to 103.70%, agreed well with the results obtained by HPLC methods, suggesting the sensor has good accuracy and high potential in practical applications.

Impact of OTAbZIP on Ochratoxin A production, mycelium growth and pathogenicity of Aspergillus westerdijkiae under water activity stress

ABSTRACT Aspergillus westerdijkiae is a major producer of ochratoxin A (OTA), a highly toxic and carcinogenic mycotoxin found in various food and feed products. A. westerdijkiae produces excessive amount of OTA under various water activity (aw) conditions that occur during food and feed storage. The biosynthetic gene clusters associated with OTA production include OTAbZIP, which plays a key role in controlling mycotoxin production in response to environmental conditions. This study explored the regulation of OTA biosynthesis in A. westerdijkiae fc-1, focusing on the OTAbZIP gene’s influence under aw stress. The mycelium growth of A. westerdijkiae fc-1 wild type and OTAbZIP mutant strains increased by 40.7% and 50.5% under high water activity (0.96 aw) respectively, at 6 days post-inoculation (dpi), indicating a stress on A. westerdijkiae fc-1. While OTAbZIP mutant did not produce OTA under both high and moderate aW conditions. The wild type produced OTA and OTA biosynthetic gene expression levels were downregulated under high (0.96 aw) and moderate (0.91 aw) water activity. The expression level of hog1 gene in OTAbZIP mutant was significantly lower than in the wild type. Pathogenicity tests revealed that deletion of OTAbZIP did not significantly affect disease infection. This study shows that deleting OTAbZIP gene greatly reduces OTA production, affecting the strain’s adaptability to water activity stress.

Study the Correlation of Ochratoxin A with a Prevalence of Breast Cancer in Al-Najaf Province, Iraq

Abstract Background: Breast cancer is a prevalent disease affecting numerous individuals worldwide. This study focuses on investigating the concentration of Ochratoxin A (OTA) in female breast cancer patients in the Al-Najaf Province. Previous research suggests a potential correlation between OTA and breast cancer. Objectives: The objective of this study was to determine the concentration of OTA and its association with breast cancer in females in the Al-Najaf Province. Materials and Methods: A total of 40 female breast cancer patients who received treatment at The National Educational Oncology Hospital in the Al-Najaf Province from April to August 2022 were included in this study. Additionally, 20 healthy females served as the control group. Serum samples were collected in sterile tubes (Eppendorf tube) and analyzed using high-performance liquid chromatography (HPLC) to quantitatively diagnose the OTA ratio. Patient information, including age, was recorded in a data sheet. Results: The results demonstrated a significant increase (P &lt; 0.05) in OTA concentrations among the total number of breast cancer patients compared to the control group. The concentrations of OTA were measured at 0.2720 and 0.2186 ng/mL in breast cancer patients, indicating a higher OTA level in this group. Conclusion: This study provides evidence of a significant association between OTA concentration and breast cancer in female patients from the Al-Najaf Province. These findings contribute to the understanding of the role of OTA in breast cancer development. Further research is warranted to explore the mechanisms underlying this relationship and potentially develop targeted interventions.

Glucose and HODEs regulate Aspergillus ochraceus quorum sensing through the GprC-AcyA pathway

Aspergillus ochraceus is the traditional ochratoxin A (OTA)-producing fungus with density-dependent behaviors, which is known as quorum sensing (QS) that is mediated by signaling molecules. Individual cells trend to adapt environmental changes in a “whole” flora through communications, allowing fungus to occupy an important ecological niche. Signals perception, transmission, and feedback are all rely on a signal network that constituted by membrane receptors and intracellular effectors. However, the interference of density information in signal transduction, which regulates most life activities of Aspergillus, have yet to be elucidated. Here we show that the G protein-coupled receptor (GPCR) to cAMP pathway is responsible for transmitting density information, and regulates the key point in life cycle of A. ochraceus. Firstly, the quorum sensing phenomenon of A. ochraceus is confirmed, and identified the density threshold is 103 spores/mL, which represents the low density that produces the most OTA in a series quorum density. Moreover, the GprC that classified as sugar sensor, and intracellular adenylate cyclase (AcyA)-cAMP-PKA pathway that in response to ligands glucose and HODEs are verified. Furthermore, GprC and AcyA regulate the primary metabolism as well as secondary metabolism, and further affects the growth of A. ochraceus during the entire life cycle. These studies highlight a crucial G protein signaling pathway for cell communication that is mediated by carbohydrate and oxylipins, and clarified a comprehensive effect of fungal development, which include the direct gene regulation and indirect substrate or energy supply. Our work revealed more signal molecules that mediated density information and connected effects on important adaptive behaviors of Aspergillus ochraceus, hoping to achieve comprehensive prevention and control of mycotoxin pollution from interrupting cell communication.