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Research on Pathogenic Fungi and Mycotoxins in China

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A special issue of Toxins (ISSN 2072-6651). This special issue belongs to the section "Mycotoxins".

Deadline for manuscript submissions: closed (31 January 2022) | Viewed by 25426

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Special Issue Editors

Prof. Dr. Shihua Wang

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Guest Editor

Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology of Education Ministry, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Interests: fungi; A. flavus; secondary metabolite; mycotoxins; biosynthetic pathway; antibody; detection; regulation; control; post-translation modification
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Yang Liu

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Guest Editor

School of Food Science and Engineering, Foshan University, Foshan 528231, China
Interests: fungi toxins; A. flavus; prevention, control and detoxification
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Qi Zhang

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Guest Editor

National Reference Lab for Agricultural Testing (Biotoxin), Key Lab of Detection for Mycotoxin, Ministry of Agriculture and Rural Affairs PRC, Lab of Quality & Safety Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture and Rural Affairs PRC, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, China
Interests: fluorescence immunosensor; functional biomaterials and nanomaterials for capture; analysis; diagnosis; warning; prevention of and reduction in toxins
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The pathogenic fungi are abundant and diverse in China, which not only directly affect the health of plants, animals, and humans but can also produce mycotoxins in China. To control pathogenic fungi, Chinese researchers have made great progress in the species identification, construction of detection methods, and pathogenic mechanism of these fungi. Importantly, mycotoxins, as the most important hazards, threaten the sustainable development of food security and feed safety. Research on the biosynthesis and regulation of mycotoxins is very important for controlling mycotoxins. Detection, especially on-site rapid detection, is a means to discover the risk of mycotoxins. Prediction and early warning is a critical way to prevent and control pollution and hazards of mycotoxins. Meanwhile, detoxification is the ideal way to reduce the hazardous effect of mycotoxins. Hence, this topic is very welcome, but not limited to the research progress of mycotoxin production, biosynthesis, and regulation of mechanisms of secondary metabolites, control of mycotoxins, detection, prediction and early warning, detoxification, and prevention and control for pathogenic fungi and mycotoxins in China.

Prof. Dr. Shihua Wang
Prof. Dr. Yang Liu
Dr. Qi Zhang
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a double-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Toxins is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

pathogenic fungi
mycotoxins
biosynthesis mechanism
detection
prediction and warning
detoxification
prevention and control
pathogenic mechanism

Related Special Issues

Research on Pathogenic Fungi and Mycotoxins in China (Volume II) in Toxins (11 articles)
Aspergillus flavus and Aflatoxins (Volume III) in Toxins (2 articles)

Published Papers (10 papers)

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Research

14 pages, 7290 KiB

Open AccessArticle

Detection of αB-Conotoxin VxXXIVA (αB-CTX) by ic-ELISA Based on an Epitope-Specific Monoclonal Antibody

by Hengkun Tang, Haimei Liu, Yehong Gao, Rui Chen, Mingke Dong, Sumei Ling, Rongzhi Wang and Shihua Wang

Toxins 2022, 14(3), 166; https://doi.org/10.3390/toxins14030166 - 23 Feb 2022

Cited by 2 | Viewed by 2002

Abstract

In view of the toxicological hazard and important applications in analgesics and cancer chemotherapeutics of αB-CTX, it is urgent to develop an accurate, effective and feasible immunoassay for the determination and analysis of αB-CTX in real samples. In this study, MBP-αB-CTX4 tandem fusion protein was used as an immunogen to elicit a strong immune response, and a hybridoma cell 5E4 secreting IgG2b against αB-CTX was successfully screened by hybridoma technology. The affinity of the purified 5E4 monoclonal antibody (mAb) was 1.02 × 10⁸ L/mol, which showed high affinity and specificity to αB-CTX. Epitope 1 of αB-CTX is the major binding region for 5E4 mAb recongnization, and two amino acid residues (14L and 15F) in αB-CTX were critical sites for the interaction between αB-CTX and 5E4 mAb. Indirect competitive ELISA (ic-ELISA) based on 5E4 mAb was developed to detect and analyze αB-CTX in real samples, and the linear range of ic-ELISA to αB-CTX was 117–3798 ng/mL, with a limit of detection (LOD) of 81 ng/mL. All the above results indicated that the developed ic-ELISA had high accuracy and repeatability, and it could be applied for αB-CTX detection and drug analysis in real samples. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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14 pages, 4754 KiB

Open AccessArticle

Toxic Effects of Mycotoxin Fumonisin B1 at Six Different Doses on Female BALB/c Mice

by Zhiwei Chen, Fan Zhang, Lin Jiang, Zihan Chen and Hua Sun

Toxins 2022, 14(1), 21; https://doi.org/10.3390/toxins14010021 - 29 Dec 2021

Cited by 9 | Viewed by 1961

Abstract

Background: Fumonisin B1 (FB1) is one of the most common mycotoxins contaminating feed and food. Although regulatory limits about fumonisins have been established in some countries, it is still very important to conduct research on lower doses of FB1 to determine the tolerance limits. The aim of this study was to investigate the effects of different concentrations of FB1, provide further evidence about the toxic doses- and exposure time-associated influence of FB1 on mice, especially low levels of FB1 for long-term exposure. Methods: Female BALB/c mice were treated intragastrically (i.g.) with fumonisin B1 (FB1) solutions (0 mg/kg body weight (BW), 0.018 mg/kg BW, 0.054 mg/kg BW, 0.162 mg/kg BW, 0.486 mg/kg BW, 1.458 mg/kg BW and 4.374 mg/kg BW) once a day for 8 weeks to obtain dose- and time-dependent effects on body and organ weights, hematology, blood chemical parameters and liver and kidney histopathology. Results: After the long-term administration of FB1, the body weights of the mice tended to decrease. Over time, FB1 first increased the relative spleen weight, then increased the relative kidney weight, and finally increased the relative liver weight. The mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), hemoglobin (HGB), white blood cells (WBC), platelets (PLT), and mean platelet volume (MPV) were significantly elevated after treatment with FB1 for 8 weeks. Moreover, exposure time-dependent responses were found for aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) level, which were coupled with hepatic histopathological findings, necroinflammation and vacuolar degeneration and detrital necrosis. Linear dose response was also found for liver histopathology, in which, even the minimum dose of FB1 exposure also caused changes. Renal alterations were moderate compared to hepatic alterations. Conclusion: In conclusion, we demonstrated the systemic toxic effects of different doses of FB1 in female BALB/c mice at different times. Our data indicated that the effects observed in this study at the lowest dose tested are discussed in relation to the currently established provisional maximum tolerable daily intake (PMTDI) for fumonisins. This study suggested that recommendations for the concentration of FB1 in animals and humans are not sufficiently protective and that regulatory doses should be modified to better protect animal and human health. The toxicity of FB1 needs more attention. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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11 pages, 2031 KiB

Open AccessArticle

Development of Generic Immuno-Magnetic Bead-Based Enzyme-Linked Immunoassay for Ustiloxins in Rice Coupled with Enrichment

by Yi Huang, Xiaoqian Tang, Lu Zheng, Junbin Huang, Qi Zhang and Hao Liu

Toxins 2021, 13(12), 907; https://doi.org/10.3390/toxins13120907 - 17 Dec 2021

Cited by 7 | Viewed by 2408

Abstract

Ustiloxins are a group of mycotoxins produced by rice false smut pathogen. Previous studies have shown that the false smut balls contain six types of ustiloxins, and these toxins are toxic to living organisms. Thus, immunoassay for on-site monitoring of ustiloxins in rice is urgently required. The current immunoassays are only for detecting single ustiloxin, and they cannot meet the demand for synchronous and rapid detection of the group toxins. Therefore, this study designed and synthesized a generic antigen with ustiloxin G as material based on the common structure of the mycotoxins. Ustiloxin G was conjugated to two carrier proteins including bovine serum albumin (BSA) and ovalbvmin (OVA) by carbon diimide method. The mice were immunized with ustiloxin-G-BSA to generate the antibody serum, which was further purified to obtain the generic antibody against ustiloxins. The conjugated ustiloxin G-OVA and generic antibodies were used for establishing the enzyme-linked immunosorbent assay (ELISA) for ustiloxin detection and optimizing experiment conditions. The characterization of the antibody showed that the semi-inhibitory concentrations (IC₅₀) of ustiloxin A, B, and G were 0.53, 0.34, and 0.06 µg/mL, respectively, and that their corresponding cross-reactivities were 11.9%, 18.4%, and 100%, respectively. To increase ELISA detection efficiency, generic antibody was combined with magnetic beads to obtain sensitive and class-specific immune-magnetic beads. Based on these immuno-magnetic beads, a high-efficiency enzyme-linked immunoassay method was developed for ustiloxin detection, whose sensitivity to ustiloxin A, B, and G was improved to 0.15 µg/mL, 0.14 µg/mL, and 0.04 µg/mL, respectively. The method accuracy was evaluated by spiking ustiloxin G as standard, and the spiked samples were tested by the immune-magnetic bead-based ELISA. The result showed the ustiloxin G recoveries ranged from 101.9% to 116.4% and were accepted by a standard HPLC method, indicating that our developed method would be promising for on-site monitoring of ustiloxins in rice. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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13 pages, 3167 KiB

Open AccessArticle

aflN Is Involved in the Biosynthesis of Aflatoxin and Conidiation in Aspergillus flavus

by Kunzhi Jia, Lijuan Yan, Yipu Jia, Shuting Xu, Zhaoqi Yan and Shihua Wang

Toxins 2021, 13(11), 831; https://doi.org/10.3390/toxins13110831 - 22 Nov 2021

Cited by 7 | Viewed by 2038

Abstract

Aspergillus flavus poses a threat to society economy and public health due to aflatoxin production. aflN is a gene located in the aflatoxin gene cluster, but the function of AflN is undefined in Aspergillus flavus. In this study, aflN is knocked out and overexpressed to study the function of AflN. The results indicated that the loss of AflN leads to the defect of aflatoxin biosynthesis. AflN is also found to play a role in conidiation but not hyphal growth and sclerotia development. Moreover, AlfN is related to the response to environmental oxidative stress and intracellular levels of reactive oxygen species. At last, AflN is involved in the pathogenicity of Aspergillus flavus to host. These results suggested that AflN played important roles in aflatoxin biosynthesis, conidiation and reactive oxygen species generation in Aspergillus flavus, which will be helpful for the understanding of aflN function, and will be beneficial to the prevention and control of Aspergillus flavus and aflatoxins contamination. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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16 pages, 1263 KiB

Open AccessArticle

Simultaneous Detection of Seven Alternaria Toxins in Mixed Fruit Puree by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry Coupled with a Modified QuEChERS

by Jiali Xing, Zigeng Zhang, Ruihang Zheng, Xiaorong Xu, Lingyan Mao, Jingping Lu, Jian Shen, Xianjun Dai and Zhenfeng Yang

Toxins 2021, 13(11), 808; https://doi.org/10.3390/toxins13110808 - 16 Nov 2021

Cited by 5 | Viewed by 2097

Abstract

The presence of Alternaria toxins (ATs) in fruit purees may cause potential harm to the life and health of consumers. As time passes, ATs have become the key detection objects in this kind of food. Based on this, a novel and rapid method was established in this paper for the simultaneous detection of seven ATS (tenuazonic acid, alternariol, alternariol monomethyl ether, altenuene, tentoxin, altenusin, and altertoxin I) in mixed fruit purees using ultra-high performance liquid chromatography-tandem mass spectrometry. The sample was prepared using the modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) method to complete the extraction and clean-up steps in one procedure. In this QuEChERS method, sample was extracted with water and acetonitrile (1.5% formic acid), then salted out with NaCl, separated on an ACQUITY UPLC BEH C₁₈ with gradient elution by using acetonitrile and 0.1% formic acid aqueous as eluent, and detected by UPLC-MS/MS under positive (ESI⁺) and negative (ESI⁻) electrospray ionization and MRM models. Results showed that the seven ATs exhibited a good linearity in the concentration range of 0.5–200 ng/mL with R² > 0.9925, and the limits of detection (LODs) of the instrument were in the range of 0.18–0.53 μg/kg. The average recoveries ranged from 79.5% to 106.7%, with the relative standard deviations (RSDs) no more than 9.78% at spiked levels of 5, 10, and 20 μg/kg for seven ATs. The established method was applied to the determination and analysis of the seven ATs in 80 mixed fruit puree samples. The results showed that ATs were detected in 31 of the 80 samples, and the content of ATs ranged from 1.32 μg/kg to 54.89 μg/kg. Moreover, the content of TeA was the highest in the detected samples (23.32–54.89 μg/kg), while the detection rate of Ten (24/31 samples) was higher than the other ATs. Furthermore, the other five ATs had similar and lower levels of contamination. The method established in this paper is accurate, rapid, simple, sensitive, repeatable, and stable, and can be used for the practical determination of seven ATs in fruit puree or other similar samples. Moreover, this method could provide theory foundation for the establishment of limit standard of ATs and provide a reference for the development of similar detection standard methods in the future. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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20 pages, 1888 KiB

Open AccessArticle

Experimental Study on the Status of Maize Mycotoxin Production in Farmers’ Grain Storage Silos in Northeastern China

by Jinsong Zhang, Yan Xu, Taogang Hu, Changpo Sun and Wenfu Wu

Toxins 2021, 13(11), 741; https://doi.org/10.3390/toxins13110741 - 20 Oct 2021

Cited by 2 | Viewed by 2118

Abstract

The scientific rationality of farmers’ grain storage technology and equipment is crucial for the biosecurity of grain in the main grain-producing areas represented by Northeast China. In this paper, four farmer grain storage mock silos of different widths were used as a means to track an experimental cycle of grain storage. The absolute water potential of corn in all four silos at the beginning of the experiment was greater than the absolute water potential of air, prompting moisture migration from the grain interior to the air and down to about 14%. Moisture was influenced by wind direction, and moisture decreased faster with better ventilation on both sides of the grain silos. Therefore, grain silo width has a significant effect on the drying effect under naturally ventilated conditions of maize ears. This research focused on the determination and assessment of mycotoxin contamination under farmers’ storage grain conditions and analyzed the effect of silo structure on the distribution of mycotoxin contamination. When the width was too large, areas of high mycotoxin infection existed in the middle of the grain silo, and ventilation and tipping could be used to reduce the risk of toxin production. This study proved that reasonable farmer grain storage techniques and devices in Northeast China can effectively protect grain from mycotoxin contamination. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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14 pages, 6306 KiB

Open AccessArticle

Zearalenone Affect the Intestinal Villi Associated with the Distribution and the Expression of Ghrelin and Proliferating Cell Nuclear Antigen in Weaned Gilts

by Quanwei Zhang, Libo Huang, Bo Leng, Yang Li, Ning Jiao, Shuzhen Jiang, Weiren Yang and Xuejun Yuan

Toxins 2021, 13(10), 736; https://doi.org/10.3390/toxins13100736 - 19 Oct 2021

Cited by 8 | Viewed by 4688

Abstract

This study explored and investigated how zearalenone (ZEA) affects the morphology of small intestine and the distribution and expression of ghrelin and proliferating cell nuclear antigen (PCNA) in the small intestine of weaned gilts. A total of 20 weaned gilts (42-day-old, D × L × Y, weighing 12.84 ± 0.26 kg) were divided into the control and ZEA groups (ZEA at 1.04 mg/kg in diet) in a 35-d study. Histological observations of the small intestines revealed that villus injuries of the duodenum, jejunum and ileum, such as atrophy, retardation and branching dysfunction, were observed in the ZEA treatment. The villi branch of the ileum in the ZEA group was obviously decreased compared to that of the ileum, jejunum and duodenum, and the number of lymphoid nodules of the ileum was increased. Additionally, the effect of ZEA (1.04 mg/kg) was decreased by the immunoreactivity and distribution of ghrelin and PCNA in the duodenal and jejunal mucosal epithelial cells. Interestingly, ZEA increased the immunoreactivity of ghrelin in the ileal mucosal epithelial cells and decreased the immunoreactivity expression of PCNA in the gland epithelium of the small intestine. In conclusion, ZEA (1.04 mg/kg) had adverse effects on the development and the absorptive capacity of the villi of the intestines; yet, the small intestine could resist or ameliorate the adverse effects of ZEA by changing the autocrine of ghrelin in intestinal epithelial cells. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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15 pages, 3368 KiB

Open AccessArticle

The Effects of Zearalenone on the Localization and Expression of Reproductive Hormones in the Ovaries of Weaned Gilts

by Boyang Wan, Xuejun Yuan, Weiren Yang, Ning Jiao, Yang Li, Faxiao Liu, Mei Liu, Zaibin Yang, Libo Huang and Shuzhen Jiang

Toxins 2021, 13(9), 626; https://doi.org/10.3390/toxins13090626 - 7 Sep 2021

Cited by 12 | Viewed by 2485

Abstract

This study aims to investigate the effects of zearalenone (ZEA) on the localizations and expressions of follicle stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), gonadotropin releasing hormone (GnRH) and gonadotropin releasing hormone receptor (GnRHR) in the ovaries of weaned gilts. Twenty 42-day-old weaned gilts were randomly allocated into two groups, and treated with a control diet and a ZEA-contaminated diet (ZEA 1.04 mg/kg), respectively. After 7-day adjustment, gilts were fed individually for 35 days and euthanized for blood and ovarian samples collection before morning feeding on the 36th day. Serum hormones of E₂, PRG, FSH, LH and GnRH were determined using radioimmunoassay kits. The ovaries were collected for relative mRNA and protein expression, and immunohistochemical analysis of FSHR, LHR, GnRH and GnRHR. The results revealed that ZEA exposure significantly increased the final vulva area (p < 0.05), significantly elevated the serum concentrations of estradiol, follicle stimulating hormone and GnRH (p < 0.05), and markedly up-regulated the mRNA and protein expressions of FSHR, LHR, GnRH and GnRHR (p < 0.05). Besides, the results of immunohistochemistry showed that the immunoreactive substances of ovarian FSHR, LHR, GnRH and GnRHR in the gilts fed the ZEA-contaminated diet were stronger than the gilts fed the control diet. Our findings indicated that dietary ZEA (1.04 mg/kg) could cause follicular proliferation by interfering with the localization and expression of FSHR, LHR, GnRH and GnRHR, and then affect the follicular development of weaned gilts. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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14 pages, 3851 KiB

Open AccessArticle

Response of Fecal Bacterial Flora to the Exposure of Fumonisin B1 in BALB/c Mice

by Fan Zhang, Zhiwei Chen, Lin Jiang, Zihan Chen and Hua Sun

Toxins 2021, 13(9), 612; https://doi.org/10.3390/toxins13090612 - 31 Aug 2021

Cited by 3 | Viewed by 2145

Abstract

Fumonisins are a kind of mycotoxin that has harmful influence on the health of humans and animals. Although some research studies associated with fumonisins have been reported, the regulatory limits of fumonisins are imperfect, and the effects of fumonisins on fecal bacterial flora of mice have not been suggested. In this study, in order to investigate the effects of fumonisin B1 (FB1) on fecal bacterial flora, BALB/c mice were randomly divided into seven groups, which were fed intragastrically with 0 mg/kg, 0.018 mg/kg, 0.054 mg/kg, 0.162 mg/kg, 0.486 mg/kg, 1.458 mg/kg and 4.374 mg/kg of FB1 solutions, once a day for 8 weeks. Subsequently, feces were collected for analysis of microflora. The V3-V4 16S rRNA of fecal bacterial flora was sequenced using the Illumina MiSeq platform. The results revealed that fecal bacterial flora of mice treated with FB1 presented high diversity. Additionally, the composition of fecal bacterial flora of FB1 exposure groups showed marked differences from that of the control group, especially for the genus types including Alloprevotella, Prevotellaceae_NK3B31_group, Rikenellaceae_RC9_gut_group, Parabacteroides and phylum types including Cyanobacteria. In conclusion, our data indicate that FB1 alters the diversity and composition of fecal microbiota in mice. Moreover, the minimum dose of FB1 exposure also causes changes in fecal microbiota to some extent. This study is the first to focus on the dose-related effect of FB1 exposure on fecal microbiota in rodent animals and gives references to the regulatory doses of fumonisins for better protection of human and animal health. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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16 pages, 3849 KiB

Open AccessArticle

The Regulatory Mechanism of Water Activities on Aflatoxins Biosynthesis and Conidia Development, and Transcription Factor AtfB Is Involved in This Regulation

by Longxue Ma, Xu Li, Xiaoyun Ma, Qiang Yu, Xiaohua Yu, Yang Liu, Chengrong Nie, Yinglong Zhang and Fuguo Xing

Toxins 2021, 13(6), 431; https://doi.org/10.3390/toxins13060431 - 21 Jun 2021

Cited by 8 | Viewed by 2233

Abstract

Peanuts are frequently infected by Aspergillus strains and then contaminated by aflatoxins (AF), which brings out economic losses and health risks. AF production is affected by diverse environmental factors, especially water activity (a_w). In this study, A. flavus was inoculated into peanuts with different a_w (0.90, 0.95, and 0.99). Both AFB₁ yield and conidia production showed the highest level in a_w 0.90 treatment. Transcriptional level analyses indicated that AF biosynthesis genes, especially the middle- and later-stage genes, were significantly up-regulated in a_w 0.90 than a_w 0.95 and 0.99. AtfB could be the pivotal regulator response to a_w variations, and could further regulate downstream genes, especially AF biosynthesis genes. The expressions of conidia genes and relevant regulators were also more up-regulated at a_w 0.90 than a_w 0.95 and 0.99, suggesting that the relative lower a_w could increase A. flavus conidia development. Furthermore, transcription factors involved in sexual development and nitrogen metabolism were also modulated by different a_w. This research partly clarified the regulatory mechanism of a_w on AF biosynthesis and A. flavus development and it would supply some advice for AF prevention in food storage. Full article

(This article belongs to the Special Issue Research on Pathogenic Fungi and Mycotoxins in China)

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