Aflatoxin B1 contamination reduces the saponins content and anti-osteoporosis efficacy of the traditional medicine Radix Dipsaci.

ETHNOPHARMACOLOGICAL RELEVANCE: The Radix Dipsaci, a traditional Chinese medicine with a history spanning over 2,000 years in China, is widely recognized for its hepatorenal tonic properties, musculoskeletal fortifying effects, fracture healing capabilities, and its frequent application in the treatment of osteoporosis. Like many traditional Chinese herbal medicines, preparations from Radix Dipsaci are at risk of contamination by harmful mycotoxins such as aflatoxin B1. AIMS OF THE STUDY: This study aims to evaluate the impact of aflatoxin B1 contamination on Radix Dipsaci in terms of changes in quality, efficacy of anti-osteoporosis and hepatorenal toxicity. MATERIALS AND METHODS: The contamination rates and levels of major mycotoxins were determined in 45 batches of Radix Dipsaci samples using UPLC-MS/MS analysis. The total saponin content and the levels of akebia saponin D in Radix Dipsaci and its decoctions were evaluated through high-performance liquid chromatography (HPLC) analysis. Differences in secondary metabolites between samples without any mycotoxin contamination (N-RD) and those contaminated solely by aflatoxin B1 (AFB1-RD) were compared using metabolomics sequencing and analysis. The anti-osteoporotic efficacy of Radix Dipsaci contaminated with aflatoxin B1 was assessed in a murine model of retinoic acid-induced osteoporosis by quantifying bone mineral content and bone mineral density using dual-energy X-ray absorptiometry. Additionally, the hepatorenal toxicity of Radix Dipsaci contaminated with aflatoxin B1 was evaluated using hematoxylin-eosin staining and enzyme-linked immunosorbent assay (ELISA). RESULTS: The results indicated that aflatoxin B1 (AFB1) was the most frequently detected mycotoxin, found in 37.7% of the Radix Dipsaci samples. AFB1 contamination significantly altered the secondary metabolites of Radix Dipsaci. Specifically, there was a notable decrease in the levels of total saponins and akebia saponin D in the AFB1-contaminated samples, which exhibited a negative correlation with the levels of AFB1 contamination. However, the administration of a water decoction from AFB1-contaminated Radix Dipsaci did not result in significant improvements in bone mineral density, bone mineral salt content, the trabecular number, trabecular area, proportion of trabecular bone volume/tissue volume and trabecular separation in an osteoporosis mouse model. Additionally, we observed that approximately 16.04% of AFB1 could migrate from the raw herbs into the decoction, leading to hepatocyte and kidney cell damage, as well as increased levels of the oxidative stress molecule malondialdehyde and pro-inflammatory cytokines in the liver and kidney tissues of the osteoporosis model mice. CONCLUSION: In summary, Radix Dipsaci is highly susceptible to mycotoxin contamination, particularly aflatoxin B1. The contamination of Radix Dipsaci with AFB1 not only impacts their saponin content and anti-osteoporosis effect but also induces hepatotoxicity and nephrotoxicity.

Sociodemographic and dietary predictors of maternal and placental mycoestrogen concentrations in a US pregnancy cohort.

BACKGROUND: Zearalenone (ZEN) is a mycotoxin contaminating grains and processed foods. ZEN alters nuclear estrogen receptor α/ß signaling earning its designation as a mycoestrogen. Experimental evidence demonstrates that mycoestrogen exposure during pregnancy is associated with altered maternal sex steroid hormones, changes in placental size, and decreases in fetal weight and length. While mycoestrogens have been detected in human biospecimens worldwide, exposure assessment of ZEN in US populations, particularly during pregnancy, is lacking. OBJECTIVE: To characterize urinary and placental concentrations of ZEN and its metabolites in healthy US pregnant people and examine demographic, perinatal, and dietary predictors of exposure. METHODS: Urine samples were collected in each trimester from pregnant participants in the UPSIDE study and placenta samples were collected at delivery (Rochester, NY, n = 317). We used high performance liquid chromatography and high-resolution tandem mass spectrometry to measure total urinary (ng/ml) and placental mycoestrogens (ng/g). Using linear regression and linear mixed effect models, we examined associations between mycoestrogen concentrations and demographic, perinatal, and dietary factors (Healthy Eating Index [HEI], ultra-processed food [UPF] consumption). RESULTS: Mycoestrogens were detected in 97% of urines (median 0.323 ng/ml) and 84% of placentas (median 0.012 ng/g). Stability of urinary mycoestrogens across pregnancy was low (ICC: 0.16-0.22) and did not correlate with placental levels. In adjusted models, parity (multiparous) and pre-pregnancy BMI (higher) predicted higher urinary concentrations. Birth season (fall) corresponded with higher placental mycoestrogens. Dietary analyses indicated that higher HEI (healthier diets) predicted lower exposure (e.g., Σmycoestrogens %∆ -2.03; 95%CI -3.23, -0.81) and higher percent calories from UPF predicted higher exposure (e.g., Σmycoestrogens %∆ 1.26; 95%CI 0.29, 2.24). IMPACT: The mycotoxin, zearalenone (ZEN), has been linked to adverse health and reproductive impacts in animal models and livestock. Despite evidence of widespread human exposure, relatively little is known about predictors of exposure. In a pregnant population, we observed that maternal ZEN concentrations varied by maternal pre-pregnancy BMI and parity. Consumption of ultra-processed foods, added sugars, and refined grains were linked to higher ZEN concentrations while healthier diets were associated with lower levels. Our research suggests disparities in exposure that are likely due to diet. Further research is needed to understand the impacts of ZEN on maternal and offspring health.

Simultaneous detection of multiple mycotoxins in Radix Dipsaci and estimation of exposure risk for consumers.

Like many traditional Chinese herbal medicines, preparations from Radix Dipsaci are at risk of contamination by harmful mycotoxins; however, there have been no reports of actual contamination. In this study, we developed an analytical method to simultaneously detect eight mycotoxins in Radix Dipsaci and estimate the exposure risk for consumers. We have developed an analytical method utilizing ultra-high performance liquid chromatography and tandem mass spectrometry to accurately determine the levels of AFB1, AFB2, AFG1, AFG2, OTA, ZEN, T-2 and ST mycotoxins in 45 batches of Radix Dipsaci sourced from major medicinal herb markets across five regions in China. We also analyzed migration of mycotoxins from the raw herbs into water decoction. Based on these results and data on human consumption of the herbal medicine, we estimated risk of exposure and acceptable exposure limits to mycotoxins in the Radix Dipsaci using the "margin of exposure (MOE)" method. Of the 45 batches of Radix Dipsaci, 48.89% contained at least one of the eight mycotoxins, 24.44% contained one, 17.78% contained two and 6.67% contained three. The most frequent mycotoxins were aflatoxin B1, present in 35.56% of batches (at 0.25-34.84 µg/kg); aflatoxin G1, 15.56% (1.99-44.05 µg/kg); and ochratoxin A, 22.22% (16.11-143.38 µg/kg). These three mycotoxins transferred from the raw herb into water decoction at respective rates of 20.20%, 29.14%, and 24.80%. The 95th percentile values of the MOE risk factors for health effects of AFB1 were below 10,000 at high doses but above 10,000 at low doses of Radix Dipsaci long-term treatment. With the reduction in duration of exposure years, the MOE values of AFB1 and AFG1 gradually reverted to within the acceptable range. The mean, 50th, and 95th percentile values of the MOE risk factors for health effects of OTA exceeded 10,000 regardless of whether consumers received a low or high dose of Radix Dipsaci treatment for durations ranging from 1 to lifetime. Based on this exposure and a typical human diet, we have estimated the respective 20-year exposure limits for Radix Dipsaci to be 5.821 µg/kg, 4.035 µg/kg, and 56.073 µg/kg for the three mycotoxins under consideration. Contamination with multiple mycotoxins is frequently observed in Radix Dipsaci, and the three most prevalent contaminants have been found to leach into water decoctions, thereby posing a potential health hazard for individuals consuming this herbal preparation. This work highlights the need to monitor herbal medicines for mycotoxin contamination in order to protect consumers.

Aflatoxin B1 in compound feed and feed ingredients from Khyber Pakhtunkhwa.

A large-scale study was conducted to explore AFB1 contamination in feed samples using Thin Layer Chromatography, following an AOAC protocol. Samples were identified which were not compliant with the maximum limits for AFB1 as regulated in the United States and Pakistan. Of a total of 923 samples, 51 samples (5.5%) were not compliant according to Pakistan and 267 (28.9%) were not compliant with US-FDA standards. The overall prevalence of non-compliant samples of compound feed was 26.2% (n = 117) according to US-FDA standards, while none of the samples were non-compliant according to Pakistan standards. Among feed ingredients, the overall prevalence of non-compliant samples was 10.7% (n = 51) and 31.4% (n = 150) according to Pakistan and USFDA standards, respectively. Non-compliant feed with respect to AFB1 contamination was highly prevalent in Khyber Pakhtunkhwa, posing a serious threat to production performance and animals health.

The relationship between aflatoxin M1 and immunoglobulin levels in cows' colostrum.

The purpose of the present research was to assess the amounts of aflatoxin M1 (AFM1) and immunoglobulin (IgA, IgG, and IgM) in cow colostrum samples, as well as their relationship. The sampling involved 90 cows (54 Montofon and 36 Simmental) from 15 independent farms. An appropriate number of samples from the total mixed ration (TMR) used in feeding the cows were collected simultaneously with the colostrum samples. AFB1 in feed, AFM1, and immunoglobulin (IgA, IgG, and IgM) levels in colostrum were evaluated using the ELISA method. The AFM1 level in colostrum samples exceeded the maximum allowed values for raw milk. Ig levels were higher in Montofon-breed cows' colostrum than in Simmentals. However, it was determined that neither the breed nor the lactation number significantly affected the colostrum Ig level. It was determined that there was no linear relationship between the number of lactations and immunoglobulin levels or between AFM1 and immunoglobulin levels in colostrum. Similarly, it was determined that there was no linear relationship between the AFB1 level in the feed and the AFM1 level in the colostrum of animals consuming these feeds.

Establishment of appropriate conditions for the efficient determination of multiple mycotoxins in tea samples and assessment of their drinking risks.

Conditions were determined for rapid, convenient, and efficient determination of 16 common mycotoxins in tea samples. Mycotoxins in tea leaves and tea infusion samples were extracted using solid-liquid extraction/liquid-liquid extraction combined with ultrasonic-assisted extraction. The extraction solvent was 2-butanone/ethyl acetate (9/1 v/v) with 0.1 % formic acid. The established conditions enabled the analysis of these mycotoxins by ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) in 5.5 min. In addition, HPLC with a temperature-controlled fluorescence detector was able to simultaneously determine 8 mycotoxins with fluorescent properties in 10 min without derivatization. Aflatoxin M1 (2.15 and 3.01 µg/kg), fumonisin B2 (198.89 µg/kg), and zearalenone (87.54 µg/kg) could be detected in commercially available pu-erh tea, green tea, and black tea products, and their total transfer rates from the products to brewed tea infusions were 64.08-65.13 %, 90.59 %, and 25.99 %, respectively. The risks of drinking mycotoxins from these tea infusions mostly showed low levels of concern.

Deoxynivalenol exposure disturbs the cytoplasmic maturation in porcine oocytes.

Deoxynivalenol (DON) is a secondary metabolite of Fusarium fungi and belonged to trichothecenes, and it widely presents in various food commodities. Previous studies have highlighted its potent toxicity, adversely affecting the growth, development, and reproductive in both humans and animals. However, the potential impact of DON on porcine oocyte organelles remains elusive. In present study, we delved into the toxic effects of DON on mitochondria, endoplasmic reticulum, Golgi during the porcine oocyte maturation. Our findings revealed that DON exposure significantly impeded granulosa cell diffusion and the expulsion of the first polar body. Additionally, mitochondrial fluorescence intensity and membrane potential underwent notable alterations under DON exposure. Notably, lysosomal fluorescence intensity decreased significantly, suggesting protein degradation and potential autophagy, which was further corroborated by the enhanced fluorescence intensity of LC3. Furthermore, endoplasmic reticulum fluorescence intensity declined, and DON exposure elevated endoplasmic reticulum stress levels, evident from the upregulated expression of GRP78. Concurrently, we observed disruption in the fusiform cortex distribution of the Golgi apparatus, characterized by reduced Golgi apparatus fluorescence intensity and GM130 expression. Collectively, our results indicate that DON exposure profoundly affects the fundamental functions of porcine oocyte organelles during meiosis and maturation.

Research diversity and advances in simultaneous removal of multi-mycotoxin.

Mycotoxins are toxic secondary metabolites produced by different fungal species under specific environmental conditions. The common and regulated mycotoxins are such as deoxynivalenol (DON), zearalenone (ZEN), ochratoxin (OTA), aflatoxin B1 (AFB1), and fumonisins (FB). These mycotoxins are highly regulated in feed and food because their effects start to exert from their lowest exposures and are abundant in our common environment. However, there are other emerging mycotoxins such as apicidin, beauvericin, aurofusarin, and enniatins which are also harmful. Thus, making a total of around 500 forms of mycotoxins. The existence of mycotoxins in feed and food has a significant impact on animal and human health, which ultimately, slows down economic growth globally. According to this review, different approaches to removing multi-mycotoxin separately or simultaneously have been stated. Mostly, the review focused on the simultaneous removal of different multiple mycotoxins. This is because the current studies show a growing trend in reporting the co-existence of multiple mycotoxins in feed and food materials, however, most detoxifying approaches are for singular mycotoxins. Therefore, the physical, chemical, and biological approaches to remove multi-mycotoxin have been elucidated as well as their advantages and limitations. Furthermore, the authors give suggestions on the way forward to reduce exposure to mycotoxins and diminish their health effects in society. Lastly, the authors emphasized introducing more stringent limits for co-existing mycotoxins, especially those that have the same health effects by acting synergistically, such as AFB1 and OTA, which both act as carcinogenic agents.

Reactions of citrinin with amino compounds modelling thermal food processing.

Citrinin (CIT) is a nephrotoxic mycotoxin, produced by several species of Penicillium, Aspergillus, and Monascus. The foodstuffs most frequently contaminated with CIT include cereals, cereal products, and red yeast rice. Studies on the occurrence of CIT in food have shown that the CIT concentrations in processed cereal-based products are generally lower than in unprocessed industry cereal samples. One possible explanation is the reaction of CIT with major food components such as carbohydrates or proteins to form modified CIT. Such modified forms of CIT are then hidden from conventional analyses, but it is possible that they are converted back into the parent mycotoxin during digestion. The aim of this study is therefore to investigate reactions of CIT with food matrix during thermal processes and to gain a deeper understanding of the degradation of CIT during food processing. In this study, we could demonstrate that CIT reacts with amino compounds such as proteins, under typical food processing conditions, leading to modified forms of CIT.

The Emerging Fusarium graminearum NA3 Population Produces High Levels of Mycotoxins in Wheat and Barley.

Fusarium graminearum (Fg) is the primary causal agent of Fusarium head blight (FHB) in wheat, barley, and other small grains in North America and worldwide. FHB results in yield reduction and contaminates grain with mycotoxins that pose threats to human and livestock health. Three genetically distinct North American (NA) populations of Fg have been characterized, which are generally associated with differences in their predominant trichothecene chemotype: NA1/15-acetyl-deoxynivalenol (15-ADON), NA2/3-acetyl-deoxynivalenol (3-ADON), and NA3/3α-acetoxy, 7,15-dihydroxy-12,13-epoxytrichothec-9-ene (NX-2). Recent studies found that the NA3 population had significantly less spread on point-inoculated wheat spikes than the NA1 and NA2 populations, and NX toxins are important for Fg spread and initial infection in wheat. In this follow-up study, to compare the effect of the three populations on initial infection and mycotoxin production on different hosts, we dip-inoculated spikes of the moderately resistant wheat cultivar Alsen and the susceptible barley cultivar Voyager using five strains from each population to evaluate disease, trichothecene mycotoxin accumulation, and trichothecene production per unit of fungal biomass. In dip-inoculated wheat spikes, the NA3 population produced significantly more trichothecene per unit of fungal biomass and accumulated higher levels of trichothecene per plant biomass than the NA1 and NA2 populations, regardless of the disease levels caused by the three populations. In contrast to its critical role during wheat infection, NX toxins had no effect on barley infection. In dip-inoculated barley, the NA1 population was more infectious and caused more severe FHB symptoms than the NA2 and NA3 populations; however, the NA3 population produced significantly higher toxin per unit of fungal biomass in infected barley tissues than the NA1 population. This study provides critical information on the emerging NA3 population, which produces high levels of NX toxin and poses a potential food safety concern.

Effects of mycotoxin-producing fungi on the fitness and gut bacterial community of the soil springtail Folsomia candida.

Mycotoxin-producing fungi are widespread and their adverse effects on mammals have been investigated; however, their impacts on soil invertebrates are not fully understood. Folsomia candida is a model soil arthropod that represents an important part of the soil invertebrate community. This study investigated the consequences of F. candida grazing on mycotoxin-producing fungi Fusarium verticillioides, F. graminearum, Aspergillus ochraceus, and A. nidulans. Consuming mycotoxin-producing fungi affected the body size and reproductive ability of F. candida, and altered the gut bacterial composition, with decreased Proteobacteria and increased Actinobacteria (Microbacterium) abundances. Notably, the abundance of foodborne fungi can be detected. Furthermore, certain bacteria isolated from F. candida's gut inhibited the growth of corresponding mycotoxin-producing fungi. The gut bacteria that inhibited mycotoxin-producing fungi growth in Aspergillus groups were also associated with poor fitness parameters and larger disruption in gut microbiota. Importantly, switching back to yeast diets reversed both the fitness parameters and gut bacterial composition. Together, our study demonstrated that grazing of mycotoxin-producing fungi by F. candida resulted in reduced physiological parameters and disturbed the gut bacterial community, and those changes can be restored by switching back to yeast diets, which indicates a strong resilience of springtails to mycotoxin-producing fungi. IMPORTANCE: Mycotoxin-producing fungi are widespread in nature and raise concerns for human and livestock health. Although they share the same ecosystem, interactions between mycotoxin-producing fungi and soil arthropods are not well understood. In this study, we report an unexpected finding that the soil arthropod Folsomia candida is rather tolerant to these mycotoxin-producing fungi. F. candida can survive solely on mycotoxin-producing fungi as a food source with reduced physiological parameters. Moreover, the gut microbial community is disturbed by mycotoxin-producing fungi, and some of the bacteria isolated from F. candida's gut can inhibit the growth of corresponding fungi. Notably, the altered physiological parameters and gut microbiota are restored when a normal diet is reintroduced, suggesting F. candida's resilience to mycotoxin-producing fungi. These findings clarify the impact of toxin-producing diets on F. candida, shedding light on how organisms can build resilience to environmental stimuli.

Mycotoxin contamination and the nutritional content of corn targeted for animal feed.

Mycotoxin contaminated corn poses a risk to poultry production. Although mycotoxin regulatory guidelines are based on the hazards of individual mycotoxin contamination, feed and feed ingredients may be contaminated with multiple mycotoxins. The objective of this study was to assess mycotoxin co-contamination and its impact on the nutrient content of corn grain. Corn samples (n = 328) originating from various regions in the Southeastern U.S. were quantitatively analyzed for fumonisin (FUM), deoxynivalenol (DON), aflatoxin (AFB1) and zearalenone (ZEA) by HPLC-MS/MS. Nutritional content was analyzed by near-infrared spectroscopy, and color data were collected. All 328 samples were found to be contaminated with at least 1 mycotoxin: 100% contained FUM (19-24,680 µg/kg), 69.82% contained DON (0-9,640 µg/kg), 17.07% contained AFB1 (0-939 µg/kg), and 43.60% had detectable levels of ZEA (0-8,093.5 µg/kg). Most of the samples were contaminated with 2 or more mycotoxins, with only 18.29% of the samples containing a single mycotoxin. 38.41% of the samples had 2 mycotoxins present, 36.59 % had 3 mycotoxins, and 4.88% of the samples had all 4 tested mycotoxins present. Samples contaminated with AFB1 had significantly lower fat (P = 0.007) and lightness (P = 0.007); samples contaminated with DON had significantly higher starch (P < 0.001) and lower protein (P < 0.001). Samples contaminated with FUM had significantly higher protein (P = 0.008) and moisture (P = 0.019) and lower starch (P < 0.001). ZEA contaminated samples had significantly lower starch (P = 0.034). A correlation was observed between mycotoxin contamination and altered nutrient content in corn. This study provides further evidence that co-contamination of mycotoxins is the norm in corn, and that mycotoxin contamination correlates with impacts on the nutrient profile of feed corn.

A diverse Fusarium community is responsible for contamination of rice with a variety of Fusarium toxins.

Rice plays an important role in the daily diet in China and therefore its quality and safety have been of great concern. However, few systematic studies have investigated Fusarium community and toxins in rice grains. Here, we collected 1381 rice samples from Jiangsu Province in eastern China and found a higher frequency of zearalenone (ZEN), deoxynivalenol (DON), fumonisins (FBs), and beauvericin (BEA). The positive samples were individually contaminated with a minimum of one and a maximum of ten toxins. Fusarium was isolated and identified as the major fungus, which exhibited temporal and geographical distribution. The most prevalent species complexes within this genus were Fusarium incarnatum-equiseti species complex (FIESC), Fusarium fujikuroi species complex (FFSC), and Fusarium sambucinum species complex (FSAMSC). Nevertheless, the amplicon sequence analysis revealed a low relative abundance of Fusarium in the rice panicles, and the fungal community exhibited an irregular change along with the symptom's emergence. In vitro toxigenic profiles of Fusarium strains showed significant complexity and specificity depending on the type and content. FIESC strains were non-pathogenic to wheat heads and weakly pathogenic to maize ears, respectively, accumulating lower amounts of toxins than F. asiaticum and F. fujikuroi. There was no significant variation in the ability to cause panicle blight in rice among the various species tested. Our study provides detailed information about the contamination of Fusarium toxins and community in rice after harvest. This information is valuable for understanding the relationship between Fusarium and rice and for developing effective control strategies.

A Surface-Enhanced Raman Spectroscopy-Based Aptasensor for the Detection of Deoxynivalenol and T-2 Mycotoxins.

The quality of food is one of the emergent points worldwide. Many microorganisms produce toxins that are harmful for human and animal health. In particular, mycotoxins from Fusarium fungi are strictly controlled in cereals. Simple and robust biosensors are necessary for 'in field' control of the crops and processed products. Nucleic acid-based sensors (aptasensors) offer a new era of point-of-care devices with excellent stability and limits of detection for a variety of analytes. Here we report the development of a surface-enhanced Raman spectroscopy (SERS)-based aptasensor for the detection of T-2 and deoxynivalenol in wheat grains. The aptasensor was able to detect as low as 0.17% of pathogen fungi in the wheat grains. The portable devices, inexpensive SERS substrate, and short analysis time encourage further implementation of the aptasensors outside of highly equipped laboratories.

Anti-Aflatoxigenic Burkholderia contaminans BC11-1 Exhibits Mycotoxin Detoxification, Phosphate Solubilization, and Cytokinin Production.

The productivity and quality of agricultural crops worldwide are adversely affected by disease outbreaks and inadequate nutrient availability. Of particular concern is the potential increase in mycotoxin prevalence due to crop diseases, which poses a threat to food security. Microorganisms with multiple functions have been favored in sustainable agriculture to address such challenges. Aspergillus flavus is a prevalent aflatoxin B1 (AFB1)-producing fungus in China. Therefore, we wanted to obtain an anti-aflatoxigenic bacterium with potent mycotoxin detoxification ability and other beneficial properties. In the present study, we have isolated an anti-aflatoxigenic strain, BC11-1, of Burkholderia contaminans, from a forest rhizosphere soil sample obtained in Luzhou, Sichuan Province, China. We found that it possesses several beneficial properties, as follows: (1) a broad spectrum of antifungal activity but compatibility with Trichoderma species, which are themselves used as biocontrol agents, making it possible to use in a biocontrol mixture or individually with other biocontrol agents in an integrated management approach; (2) an exhibited mycotoxin detoxification capacity with a degradation ratio of 90% for aflatoxin B1 and 78% for zearalenone, suggesting its potential for remedial application; and (3) a high ability to solubilize phosphorus and produce cytokinin production, highlighting its potential as a biofertilizer. Overall, the diverse properties of BC11-1 render it a beneficial bacterium with excellent potential for use in plant disease protection and mycotoxin prevention and as a biofertilizer. Lastly, a pan-genomic analysis suggests that BC11-1 may possess other undiscovered biological properties, prompting further exploration of the properties of this unique strain of B. contaminans. These findings highlight the potential of using the anti-aflatoxigenic strain BC11-1 to enhance disease protection and improve soil fertility, thus contributing to food security. Given its multiple beneficial properties, BC11-1 represents a valuable microbial resource as a biocontrol agent and biofertilizer.

CRISPR/Cas12a-Based Indirect Competitive Enzyme-Linked Immunosorbent Assay for Sensitive Detection of Ochratoxin A.

The high toxicity and widespread contamination of ochratoxin A (OTA) make it urgent to develop a sensitive method to detect trace OTA in complex food matrices. Herein, an indirect competitive enzyme-linked immunosorbent assay (icELISA)-based on the CRISPR/Cas12a system is described. DNA amplicons with multiple activation sequences of the CRISPR/Cas12a system were pre-prepared to improve detection sensitivity. In the absence of OTA, streptavidin-mediated biotinylated DNA amplicons were captured by the biotinylated secondary antibody on the microplate. The captured DNA amplicons activated the CRISPR/Cas12a system, which thereby effectively cleaved the reporter DNA, producing strong fluorescence. The presence of OTA led to a decrease in DNA amplicons on the microplate, resulting in a decrease in activated Cas12a and ultimately a drop in fluorescence intensity. OTA in food matrices at nanogram per milliliter levels can be detected. Therefore, the new method has great potential in monitoring OTA.

Bacillus velezensis CL197: a zearalenone detoxifying strain isolated from wheat with potential to be used in animal production.

Zearalenone (ZEA) is a mycotoxin produced by Fusarium species, and cause contamination of food and feed, with impacts in animal production and in food production chain. Effective detoxifying methods, such as biodegradation, are therefore required. This study aimed to isolate microorganisms and screen ZEA detoxifying strains. As a result, 197 microorganisms were isolated, and six were initially selected after colorimetric screening. ZEA (1 µg/mL) was added to culture media, and after 24 h, all six microorganisms were able to degrade ZEA, without the formation of α-ZOL. One isolate eliminated ~ 99% of ZEA and was identified as Bacillus velezensis CL197. ZEA metabolites produced by the bacteria were evaluated, and no metabolites with greater or similar toxicity than ZEA were detected. This strain was applied to swine in vitro digestion, and up to 64% of ZEA was degraded. B. velezensis CL197 significantly degraded ZEA, demonstrating potential to be used as a detoxifying agent in the food production chain as a biocontrol agent.

New application of a dye-decolorizing peroxidase immobilized on magnetic nanoparticles for efficient simultaneous degradation of two mycotoxins.

Nowadays the enzymatic approaches are the most promising strategies for mycotoxins detoxification in food stuffs. Herein, the dye-decolorizing peroxidase RhDypB from Rhodococcus jostii was studied for its ability to degrade two mycotoxins in both free and the immobilized enzyme forms. This enzyme was recombinantly expressed and purified, while Fe3O4 nanoparticles were prepared and modified with chitosan as the immobilization carrier. The immobilized enzyme Fe3O4@CS@RhDypB demonstrated degradation rate of 85.61 % toward aflatoxin B1, while it was firstly found to be able to degrade zearalenone with the rate of 86.52 %, at pH 4.0 on 30 °C. The degradation products were identified as aflatoxin Q1 and 15-OH-ZEN respectively. After 5 cycles of reuse, Fe3O4@CS@RhDypB still exhibited degradation rates of 38.50 % and 49.76 % toward the mycotoxins, indicating its high reusability. Moreover, Fe3O4@CS@RhDypB exhibited excellent stability after 10 days of storage. This work identified potential applications of nanoparticle-immobilized enzyme for biodegradation of mycotoxins in food industry.

Endoplasmic reticulum stress is involved in mycotoxin zearalenone induced inflammatory response, proliferation, and apoptosis in goat endometrial stromal cells.

Zearalenone (ZEA) is an estrogen-like mycotoxin and is considered a secondary metabolite produced by Fusarium fungi, which are widely found in the surrounding environment. ZEA has been found to cause reproductive dysfunction in female and male animals, but the underlying mechanism remains unclear. Therefore, this study examined cell proliferation, cell apoptosis, autophagy protein expression, and some inflammatory cytokines such as IL-1ß and IL-8 of goat endometrial stromal cells (ESCs) induced by different concentrations (0, 15, 30, 60, and 90 µM) of ZEA. The apoptosis rate was detected by flow cytometry. Western Blot and ELISA assay were used to identify the ER stress signaling pathway and some inflammatory cytokines. Our results revealed that ZEA induced cell proliferation and inhibited cell apoptosis at low and middle concentrations, while at high concentrations of ZEA, cell apoptosis was induced in ESCs. Additionally, ZEA induced the ER stress protein markers such as ATF6, IRE1α, EIF2α, and ATF4. LC3 as a marker of autophagy was up-regulated at all concentrations of ZEA. Moreover, IL-1ß and IL-8 showed down-regulation at a low concentration of ZEA, but middle and high concentrations showed up-regulation. In the present study, Knockdown ERN1 can inhibit autophagy and the main markers of ER stress. These results suggest that the IRE1 pathway can reduce apoptosis protein markers, down activate IRE1, and unfolded protein response branches such as ATF6 and LC3 in ESCs. Additionally, IL-1ß and IL-8 achieve up-regulation under knockdown IRE1, which can block ER stress markers.