Morphological and molecular response of small intestine to lactulose and hydrogen-rich water in female piglets fed Fusarium mycotoxins contaminated diet

Background: Following the intake of Fusarium mycotoxin-contaminated feed,small intestines may be exposed to high levels of toxic substances that can potentially damage intestinal functions in livestock.It is well known that Fusarium mycotoxins will lead a breakdown of the normally impeccable epithelial barrier,resulting in the development of a "leaky" gut.H2 administration with different methods has been proved definitely potentials to prevent serious intestinal diseases.The goal of this study is to investigate the roles of lactulose(LAC) and hydrogenrich water(HRW) in preventing intestinal dysfunction in piglets fed Fusarium mycotoxin-contaminated feed.Methods: A total of 24 female piglets were evenly assigned to 4 groups: negative control(NC) group,mycotoxincontaminated(MC) feed group,MC feed with LAC treatment(MC + LAC),and MC feed with HRW treatment(MC +HRW),respectively.Piglets in the NC group were fed uncontaminated control diet,while remaining piglets were fed Fusarium mycotoxin-contaminated diet.For the NC and MC groups,10 mL/kg body weight(BW) of hydrogen-free water(HFW) was orally administrated to piglets twice daily;while in the MC + LAC and MC + HRW groups,piglets were treated with the same dose of LAC solution(500 mg/kg BW) and HRW twice daily,respectively.On d 25,serum was collected and used for biochemical analysis.Intestinal tissues were sampled for morphological examination as well as relative genes and protein expression analysis.Results: Our data showed that Fusarium mycotoxins induced higher serum diamine oxidase(DAO) activities(P < 0.05),D-lactic acid levels(P < 0.01),and endotoxin status(P < 0.01),lower villus height(P < 0.01) and ratio of villus height to crypt depth(P < 0.05) in small intestine,greater apoptosis index and higher mRNA expression related to tight junctions(P < 0.05).In addition,the distribution and down-regulation of claudin-3(CLDN3) protein in the small intestinal was also observed.As expected,oral administrations of HRW and LAC were found to remarkably provide beneficial effects against Fusarium mycotoxin-induced apoptosis and intestinal leaking.Moreover,either HRW or LAC treatments were also revealed to prevent abnormal intestinal morphological changes,disintegrate tight junctions,and restore the expression and distribution of CLDN3 protein in the small intestinal mucosal layer in female piglets that were fed Fusarium mycotoxins contaminated diet.Conclusions: Our data suggest that orally administrations of HRW and LAC result in less Fusarium mycotoxininduced apoptosis and leak in the small intestine.Either HRW or LAC treatments could prevent the abnormal changes of intestinal morphology and molecular response of tight junctions as well as restore the distribution and expression of CLDN3 protein of small intestinal mucosa layer in female piglets that were fed Fusarium mycotoxins contaminated diet.

Traditional and emerging Fusarium mycotoxins disrupt homeostasis of bovine mammary cells by altering cell permeability and innate immune function

High incidence of traditional and emerging Fusarium mycotoxins in cereal grains and silages can be a potential threat to feed safety and ruminants.Inadequate biodegradation of Fusarium mycotoxins by rumen microflora following ingestion of mycotoxin-contaminated feeds can lead to their circulatory transport to target tissues such as mammary gland.The bovine udder plays a pivotal role in maintaining milk yield and composition,thus,human health.However,toxic effects of Fusarium mycotoxins on bovine mammary gland are rarely studied.In this study,the bovine mammary epithelial cell line was used as an in-vitro model of bovine mammary epithelium to investigate effects of deoxynivalenol(DON),enniatin B(ENB)and beauvericin(BEA)on bovine mammary gland homeostasis.Results indicated that exposure to DON,ENB and BEA for 48 h significantly decreased cell viability in a concentration-dependent manner(P<0.001).Exposure to DON at 0.39μmol/L and BEA at 2.5μmol/L for 48 h also decreased paracellular flux of FITC-40 kDa dextran(P<0.05),whereas none of the mycotoxins affected transepithelial electrical resistance after 48 h exposure.The qPCR was performed for assessment of expression of gene coding tight junction(TJ)proteins,toll-like receptor 4(TLR4)and cytokines after 4,24 and 48 h of exposure.DON,ENB and BEA significantly upregulated the TJ protein zonula occludens-1,whereas markedly downregulated claudin 3(P<0.05).Exposure to DON at 1.35μmol/L for 4 h significantly increased expression of occludin(P<0.01).DON,ENB and BEA significant downregulated TLR4(P<0.05).In contrast,ENB markedly increased expression of cytokines interleukin-6(IL-6)(P<0.001),tumor necrosis factorα(TNF-a)(P<0.05)and transforming growth factor-β(TGF-β)(P<0.01).BEA significantly upregulated IL-6(P<0.001)and TGF-β(P=0.01),but downregulated TNF-α(P<0.001).These results suggest that DON,ENB and BEA can disrupt mammary gland homeostasis by inducing cell death as well as altering its paracellular permeability and expression of genes involved in innate immune function.

Occurrence,toxicity,production and detection of Fusarium mycotoxin:a review

Fusarium mycotoxin contamination of both foods and feeds is an inevitable phenomenon worldwide.Deoxynivalenol,nivalenol,zearalenone,T-2 toxin and fumonisin B1 are the most studied Fusarium mycotoxins.Cocontamination of mycotoxins has also been studied frequently.Fusarium mycotoxins occur frequently in foods at very low concentrations,so there is a need to provide sensitive and reliable methods for their early detection.The present review provides insight on the types,toxicology and occurrence of Fusarium mycotoxins.It further elucidates various detection methods of mycotoxin production from Fusarium strains,with a special focus on chromatographic and immunochemical techniques.

Overview of analysis,risk assessment and control of Fusarium mycotoxins in contaminated foods

In the past decade,Fusarium mycotoxins as main biological contaminants in foods have attracted increasing attention due to their agro-economic importance across the global food chain.Their chemical structural diversities and occurance in a variety of foods make it chanllenging to simultaneously and efficiently extract them for further analysis.On the other hand,although various analytical methods have been widely applied for on-site and in-lab tests for rapid screening and/or quantitative determination,risk assessment of Fusarium mycotoxins is hardly performed due to the unavailability of affordable mycotoxin standards,as well as the lack of data of dietary consumption and food contamination.Meanwhile,there is an increasing demand for effective control of mycotoxins regarding prevention,neutralization,clearance,detoxification approaches using physico-chemical and biological means.This review summarizes advances in the analysis and control of Fusarium mycotoxins in contaminated foods,providing valuable insights and references for subsequent studies on development of analytical methodology,control strategies,as well as regulations of food safety made by policy-makers.

The Risk of <i>Fusarium</i>and Their Mycotoxins in the Food Chain

Fusarium sp. and mycotoxins of these species pose a major risk to consumer health, agriculture and the food industry. This paper is a worldwide bibliographic study on impact of Fusarium and mycotoxins on the food chain. The factors influencing the development of fungi Fusarium sp., the formation of mycotoxins and their microbiological risk on the food chain must be considered as a whole. For cereals and oilseeds before and after harvest, fungal infections and mycotoxin contamination present serious problems worldwide. This paper is an overview of the factors that include the microbiological risk and impact of Fusarium in the food chain mentioned in national and international studies. The methods and results obtained in this direction internationally are mentioned, such as: infrared spectroscopy, Raman spectrometry and hyperspectral imaging. Also, in review are presented solutions to reduce this impact on the food chain.

我国小麦赤霉病流行与化学防控现状及控病降毒关键技术理论创新与应用

概述了我国小麦赤霉病发生危害现状,深入剖析了该病流行危害的原因,回顾了近年来我国小麦赤霉病化学防控历程,综述了化学防控及其抗药性发生的现状,并总结了小麦赤霉病菌抗药性检测技术的发展过程。此外,对小麦赤霉病控病降毒关键技术理论创新与应用成果进行了分析和总结,展望了小麦赤霉病精准防控的科学用药技术方案,并对其可行性和有效性进行了阐释。旨在为小麦赤霉病的精准选药、科学用药提供重要的参考数据,保障我国的粮食生产安全和食品安全,进而推动我国农业实现可持续、高质量发展。

镰刀菌属真菌毒素在植物和病原菌互作中的研究进展

镰刀菌是世界上最重要的植物病原菌之一,可影响植物的生长发育,严重威胁全球粮食安全和生物多样性。几乎所有的镰刀菌都会产生真菌毒素,其毒素种类多、毒性强,一方面可以作为致病因子之一参与镰刀菌的致病过程,另一方面可污染粮食和饲料,进而引起人类和动物的相关病症。已有研究表明,镰刀菌侵染植物后产生的不同种类真菌毒素不仅毒害植物细胞,引起植物组织的坏死,还会加速病原菌的侵染;同时,针对病原菌产生的毒素,植物会激活防御酶并启动防御相关基因的表达,或将致病毒素转化为无毒或低毒物质并转运到胞外,或通过分泌次生代谢物直接抑制病原菌毒素的生物合成。为全面解析镰刀菌毒素在病原菌侵染植物中的作用,提高植物对病原菌的抗性,该文综述了镰刀菌属真菌毒素的种类、毒性机理以及毒素在植物和病原菌互作中的作用,并讨论了植物对真菌毒素的防御反应策略,以期为镰刀菌毒素致病机制和病原菌防治策略研究提供参考。

大麦中抑制镰刀霉菌的乳酸菌筛选及其在制麦中的应用

大麦表面存在多种微生物,其中镰刀霉菌及其分泌的真菌毒素会对制麦过程及麦芽品质产生影响。为了消除镰刀霉菌带来的不利影响,利用MRS选择性培养基从大麦表面分离筛选出一株对镰刀霉菌有明显抑制作用的菌株。根据菌株的形态学观察、生理生化试验以及16S rDNA基因序列分析鉴定,确定该菌株为植物乳杆菌(Lactiplantibacillus plantarum),命名为BY66。在制麦过程中,添加0.5 mL/kg的BY66发酵液能使禾谷镰刀霉的生物量和呕吐毒素含量分别降低56.2%和79.1%。BY66发酵液能消除禾谷镰刀霉菌在大麦发芽和麦芽质量方面产生的负面影响,与侵染大麦相比,添加0.5 mL/kg BY66发酵液后的大麦发芽率提高了31%。

镰刀菌真菌毒素的分类与研究

镰刀菌是甘蔗梢腐病的病原菌,它所产生的真菌毒素导致的病害,是当今世界上的一大生产防治难题。其中,玉米赤霉烯酮(Zearalenone),单端孢霉烯族毒素(Trichothecenes)、伏马菌素(Fumonisin)是当前研究中最受关注、影响极为广泛的三种镰刀菌毒素。本文阐述了甘蔗梢腐病病原镰刀菌所产毒素的主要种类、危害及相关研究,并探讨了未来可能的研究方向,为甘蔗真菌性病害研究提供相应的参考与借鉴。

小麦中致病真菌的分离鉴定及熏蒸对其毒力的影响

小麦受镰刀菌的感染会导致赤霉病、真菌毒素污染和减产,镰刀菌对粮食安全有着严重威胁。通过对小麦中镰刀菌株进行分离鉴定,分离出的三种菌株N1、N2、N3分别属于禾谷镰刀菌属、亚洲镰刀菌属和高秆镰刀菌属。利用产毒基因检测和真菌毒素检测对三株菌株玉米赤霉烯酮、呕吐毒素和伏马毒素的产毒情况进行分析,结果一致显示N1和N3能产生玉米赤霉烯酮和呕吐毒素,均含有PSK和Tri5两种产毒基因,而N2不产生这三种毒素。进一步通过臭氧和二氧化氯进行气体熏蒸,探究气体熏蒸对分离出的三株菌株的影响,通过观察孢子形态、菌丝体长度、菌丝形态,结果表明二氧化氯熏蒸能有效抑制菌丝生长和孢子萌发,低浓度二氧化氯(300 mg/L)处理0.5 h,可明显抑制菌丝的生长。而臭氧只能抑制孢子萌发,对菌丝的生长没有明显的抑制作用。

Tolerance of New Introgressive Hybrid and Backcross Forms Pathogenic Micromitisms (<i>Verticillium dahliae</i>Kleb and Fusarium oxysporum f.sp. vasinfectum)

The article is based on the use of experimental polyploidy method, with the introduction of new introgressive hybrid forms combining several species genotype with pathogenic Verticillium dahliae Kleb. and the effects of mycotoxins separated from the Fusarium oxysporum f.sp. vasinfectum micromicette on the yield of plant seeds. New artificial complex hypertension forms based on experimental polyploidy Verticillium dahliae Kleb. and Fusarium oxysporum f.sp. vasinfectum combine the potential of resistance to mycotoxins separated from microcrystals, making a tremendous contribution to the selection of new varieties and to the effectiveness of selection as a result of the use of genetic selective research as genetic-selective genetic-selector studies.

Preparation on the basis of Trichoderma asperellum in the system of biological protection of wheat from Fusarium ear scab

During the last century, as the area of wheat grown under advanced grain husbandry has increased worldwide, so too has the importance of Fusarium ear scab (FES) (synonym, Fusarium head blight) caused by several species of the fungus Fusarium. Yield losses due to FES can total 20%-40% and more depending on climatic conditions. During the last twenty years epidemics of FES in cereals have become chronic all over the world, including the United States and Russia. The most destructive of these were observed in 1982, 1986, 1990-1996 in USA and in the south of Russia in 1982, 1984, 1988, 1992. The harmful effect of FES is manifested not only in reduced grain yields, but also in the contamination of grains and grain products with mycotoxins, such as deoxynivalenol (DON) and its derivatives (3-alfa acetyl-DON, 15-alfa acetyl-DON), T-2 toxin and zearalenone. Standard means to control FES (cultural control methodologies, chemical pesticides, and FES resistant varieties) have little effect or are not practical and rarely reduce the accumulation of mycotoxins in grain. We have developed a new technique to reduce FES using biological preparations. The technique utilizes wheat seed pretreatment with a biofungicide “Mycol” in combination with spraying wheat plants during flowering with a yeast preparation. Technology for production of Mycol on the basis of Trichoderma asperellum strain GJS 03-35 (systematics by Samuels) has been developed. This strain shows hyperparasitic activity against a wide spectrum of plant pathogens, including Fusarium graminearum, a causative agent of FES in wheat. Experiments conducted in the United States demonstrated that spraying wheat plants during flowering with the patented yeast Cryptococcus nodaensis OH 182.9 (NRRL Y-30216) reliably reduces FES development. Tests of the Mycol preparation and the yeast OH 182.9 (EOD) have been performed on the spring wheat “Ivolga” in greenhouse conditions (the Moscow region) and on the winter wheat “Kupava” in field trials in the North Caucasian region. An isolate of F. graminearum was used to insure adequate levels of disease development in greenhouse and field experiments. FES disease severity and incidence, as well as mycotoxin accumulation in wheat grains was studied for single or combination treatments with the biological preparations. Mycol (in concentrations 0.1, 0.5, 1.0, 2.0 kg/t of seeds) was used for wheat seed pretreatment. The yeast preparation EOD (2.0×107 cfu/mL) was applied by spraying wheat plants during flowering. Chemical pesticides (Raxyl, TMTD) and a biological preparation Agat-25K were used as alternative control seed treatments. In greenhouse experiments, inoculations of heads with either biological preparation 4 h prior to inoculation with conidia of F. graminearum significantly reduced FES severity. Application with Mycol reduced DON in wheat grains by 6 to 11 fold. EOD alone or, to a lesser extent, in combination was also highly effective in reducing DON content. For treatments consisting of Mycol and EOD, 1000 grain weights were equivalent or higher than for control plants (both infected, and not infected). Wheat seeds obtained from the plants protected by these biological preparations germinated rapidly and possessed high germination rates compared to the FES control. In field trials, Mycol treatments clearly reduced FES symptoms, apparently providing an immunizing effect against FES. Mycol reduced FES severity and enhanced yield of the wheat varieties used. The effect of Mycol used at a minimum test-dose (0.1 kg/t) was not so pronounced. The greatest reduction of FES development was observed at a dose of Mycol of 1.0 kg per 1 t of seeds used in combination with EOD spraying. Experimental results support the contention that the offered technology has good prospects in controlling wheat Fusarium ear scab.

Occurrence of <i>Fusarium</i>species and associated T2-toxin in Kenyan wheat

Survey covering 120 wheat fields was conducted in three wheat-growing districts of Kenya during the 2008 cropping season to determine the incidence of Fusarium head blight (FHB) and T2-toxin contamination in grain. FHB incidence was determined as the number of blighted ears per 10m2. Information gathered included wheat production practices, rainfall and temperature data. Fungal pathogens were isolated from wheat stems, heads, straw, grains and soil and identified based on cultural and morphological characteristics. Wheat grain samples were analyzed for T2-toxin by competitive Enzyme Linked Immunosorbent Assay (ELISA). High FHB incidences of up to 88% were recorded. Fungal genera isolated included Fusarium, Epicoccum, Trichoderma, Alternaria and Penicilium. Wheat plant parts with high infection with Alternaria and Epicoccum had corresponding low levels of Fusarium spp. Whereas Fusarium spp. were the most common fungal pathogens in stems, heads and soil, Epicoccum was frequently isolated from straw and grains. Fusarium speciesisolated included F. poae, F. graminearum, F. stilboides, F. verticilloides, F. fusarioides, F. tricinctum and F. heterosporum with F. poae and F. graminearum accounting for approximately 40% of all Fusarium infections. T-2 toxin was detected in all the grain samples and varied from 3 to 22 ppb. The study showed that FHB and T2-toxin are prevalent in the study districts and the high diversity of Fusarium species implies a challenge in FHB management as well as a risk of chronic T2-toxin exposure to humans and livestock.

The Nature, Sources, Detections and Regulations of Mycotoxins That Contaminate Foods and Feeds Causing Health Hazards for Both Human and Animals

Mycotoxins are toxic secondary metabolites produced by fungus kingdom. Fungi (molds) under aerobic and optimum conditions of humidity and temperature consume nutrients for proliferation and mycotoxin production (secretion). There are seven major groups of mycotoxins produced by different species of toxigenic fungal genus. Mycotoxins production from these toxigenic fungi depends on the surrounding intrinsic and extrinsic environments. These seven mycotoxins groups that contaminate grains, foods and animal feeds are: Aflatoxins, Trichothecene, Ochratoxins, Ergot alkaloid (Ergolin), Fumonisins, Patulin, and Zearalenone. These mycotoxins are capable of causing health hazards and death for both human and animals by effecting mammalian cells, causing a number of problems in normal cell function and a wide variety of clinical symptoms of diseases. These mycotoxins are varied in their toxicity depending on the infected host (human or animal) and the host susceptibility (immunity). The major concern of food and feed industries is the contamination of food products and animal feed supplies by these mycotoxins. Worldwide Health Organization (WHO), and Food and Agriculture Organization (FAO) are responsible to regulate the acceptable (tolerable) levels of these mycotoxins in grains, food and feed supplies to ensure the safety and health for both human and animals. Understanding fungal ecology and factors that affect fungal proliferation and mycotoxins production by these toxigenic fungi in agriculture crops as raw materials for both human food and animal feed products, plus understanding the chemistry and property of these mycotoxins, methods of detection, illness symptoms, and comply with regulatory guidance established by World Health Organization (WHO)/Food and Agriculture Organization (FAO) are key factors to prevent or minimize foods/feeds contamination and the toxicity of these mycotoxins for both human and animals health, plus reducing economical loss.

粮食中新兴真菌毒素的污染现状及毒理学研究进展

真菌毒素是产毒真菌污染谷物粮食等农产品产生有毒有害的次级代谢产物,严重威胁食用和饲用产品的质量安全。较为常见的真菌毒素包括黄曲霉毒素、镰刀菌毒素脱氧雪腐镰刀烯醇、玉米赤霉烯酮等,国内外针对这些主要污染毒素制定了相应的限量标准。近年来,随着真菌毒素研究的发展和检测技术的更新,一些新型真菌毒素不断被发现。新兴毒素(Emerging toxins)是目前尚未被列入常规风险筛查和监管、也尚无相关限量标准的真菌毒素种类。目前检出频次高、污染较为严重的新兴毒素主要由镰刀菌属(Fusarium spp.)和链格孢属(Alternaria spp.)等产毒真菌产生。这些新兴真菌毒素通常与主要毒素复合污染,文章将依据现有的风险调查报告与数据,综述粮食中常见新兴镰刀菌和链格孢毒素的污染现状和毒理学研究进展,为真菌毒素的全面风险评估和管控技术研发提供参考。

陕西省小麦不同部位镰刀菌分离鉴定及毒素化学型分析

为明确陕西省小麦不同部位镰刀菌种群结构及其毒素化学型,于2022年小麦灌浆期分别从陕西省渭南市、宝鸡市、咸阳市、西安市采集小麦穗部、茎秆以及茎基部具有明显病状的样品,采用单孢分离法获得镰刀菌菌株,并使用分子生物学鉴定其种群结构及毒素化学型。结果表明,从采集的样本中共分离到156株镰刀菌菌株,其中发生在穗部的赤霉病以及发生在茎秆的秆腐病的优势病原菌均为禾谷镰刀菌(Fusarium graminearum),主要以15-ADON化学型为主;发生在茎基部的茎基腐病的优势病原菌为假禾谷镰刀菌(F.pseudograminearum),其15-ADON和3-ADON化学型占比差异不大;未检测到NIV毒素化学型的镰刀菌。以上结果说明,小麦不同部位对镰刀菌毒素化学型可能不存在选择性,小麦茎基部与穗部和茎秆的镰刀菌毒素化学型不同,原因可能是由优势病原菌不同造成的。

姜黄素介导光动力抑制禾谷镰刀菌产ZEN毒素的研究

玉米赤霉烯酮(Zearalenone,ZEN)是广泛污染玉米、小麦、大米等谷物的真菌毒素之一,主要由镰刀菌属产生。该研究选择姜黄素为光敏剂构建光动力技术(Photodynamic Inactivation,PDI),探究姜黄素浓度、光照能量和溶液pH对PDI抑制禾谷镰刀菌生长及其在玉米中产ZEN的影响。同时,从ZEN毒素含量变化和禾谷镰刀菌失活两个角度分析PDI抑制禾谷镰刀菌产ZEN的机制。结果表明,在50.00μmol/L姜黄素和43.0 J/cm^(2)光照能量作用下,ZEN含量从空白对照玉米中的1.15 mg/kg降低至光动力处理组中的0.58 mg/kg,降幅达49.56%,且继续增加姜黄素浓度能够完全抑制ZEN的生成。深入分析表明,PDI对禾谷镰刀菌的灭活作用是其抑制玉米中ZEN产生的主要原因,尤其当姜黄素浓度≥70.00μmol/L,光照能量为43.0 J/cm^(2)时,禾谷镰刀菌被完全灭活。因此,该研究证实光动力技术能够有效地杀灭禾谷镰刀菌,抑制ZEN的产生,为保障农产品质量安全提供了一种新方法。

镰刀菌毒素对断奶仔猪的抗营养效应及葡配甘露聚糖吸附剂的保护作用

选择125头始重(8.04±0.64)kg去势约×荣仔猪,采用2×2+1因子设计,研究被镰刀菌毒素自然污染的玉米以25%和50%的比例等量替换基础饲粮中的正常玉米并在此基础上加或不加葡配甘露聚糖(EGM)霉菌毒素吸附剂霉可吸TM对断奶仔猪生产性能、养分利用率的影响。结果表明:饲喂被镰刀菌毒素污染的饲粮降低了仔猪的生产性能,随霉变玉米替代比例增加,采食量和日增重降低程度增加(P<0.05或P<0.01);显著或极显著降低仔猪饲粮中有机物质、蛋白质、钙、磷的表观消化率(P<0.05或P<0.01)和蛋白质生物学价值(P<0.05),25%污染玉米饲粮对钙的表观消化率无影响(P>0.05);在被镰刀菌毒素污染饲粮中添加0.2%EGM吸附剂可使上述不良效应得到改善,尤以在25%污染玉米饲粮中添加0.2%EGM吸附剂可获得显著的保护效应。