14C‐labeled aflatoxin B1 prepared with yeastlike cultures of Aspergillus parasiticus

Abstract

A simple method was developed to produce ¹⁴C‐labeled aflatoxin B₁ by using the yeastlike phase of Aspergillus parasiticus NRRL 2999. Yeastlike cultures resulted from absence of manganese in a synthetic medium. Sodium acetate‐1‐¹⁴C had a 0.22% average incorporation; sodium acetate‐1,2‐¹⁴C, 0.70%. The average yield of labeled B₁ was 10 mg/500 ml medium with an average specific activity of either 63.3 mCi/mole (C‐l label) or 194.3 mCi/mole (C‐1, 2 label).     

The effect of deoxynivalenol on serotoninergic neurotransmitter levels in pig blood 1

Abstract

Deoxynivalenol (DON) produces two characteristic toxicological effects, decreased feed consumption (anorexia) and emesis. Both effects have been linked to increased central (CNS) serotoninergic activity. Although there has also been some indication of a peripheral involvement, the role of blood pools of serotonin and related compounds in mediating DON toxicity is not well defined. In this study, the effect of DON on plasma concentrations of serotonin (5‐hydroxytryptamine, 5HT), 5HIAA (5‐hydroxyindoleacetic acid) and tryptophan (TRP), as a reflection of an induced peripheral serotoninergic system, was investigated in swine.

Typical values for the plasma concentrations of 5HT, 5HIAA, and TRP were established in pigs. Following administration of DON, either intragastrically or intravenously, concentration changes in these substances were measured over an eight hour period. The effect of low and high toxin doses were also compared.

Analyses showed no effect on plasma levels of the compounds of interest, even at sufficient toxin doses to invoke emesis in the test animals. Any variation over the course of the study remained within acceptable control limits. These results indicated no peripheral effect by DON which could account for the increased serotoninergic activity associated with altered feeding behaviour or emesis.     

中国与欧洲禾谷镰刀菌DON毒素HPLC定量比较分析

选用4个来自中国、7个来自欧洲的代表性禾谷镰刀菌菌株,经脱氧雪腐镰刀菌烯醇(DON)毒素特异引物鉴定,确认其具有产生DON毒素的遗传物质基础后,接种于PDB培养基培养7d,从培养基上清中经硅胶柱分离、纯化DON毒素,经HPLC定量分析表明,纯化的禾谷镰刀菌DON毒素,与购自公司的DON毒素标准样品一样,其HPLC检测谱峰清晰明显,基线平稳,无干扰,保留时间为9.5min左右;供试菌株的DON毒素含量分布在0.023~1.934μg/mL之间,德国菌株F703产毒量最大,比位居第二的中国菌株5005(1.232μg/mL)高57%;其余的6个欧洲菌株中,除意大利菌株Lor9(0.128μg/mL)略低于另一个中国菌株7105(0.135μg/mL)外,均比3个中国菌株(4020、7105、8029)的产毒量大.欧洲镰刀菌产生DON毒素的能力远远大于中国菌株,这说明欧洲菌株长期在欧洲生态环境下已演变形成其特有的高毒素代谢类型,我国有必要严防欧洲禾谷镰刀菌入侵,加强对来自欧洲的禾谷类粮食及其产品的镰刀菌和毒素的检疫和监控;同时,本研究建立的毒素样品制备与HPLC检测体系可用于准确定量分析样品的DON毒素.图3表2参14     

Zearalenone screening of human breast milk from the Naples area

The mycotoxin zearalenone exhibits estrogenic and anabolic properties in several animal species, humans included. Food contamination by zearalenone is caused either by direct contamination of grains, fruits, and their based-products or by “carry-over” of mycotoxins in animal tissues, milk, and eggs after intake of contaminated feedstuff. Now, a survey on zearalenone contamination in breast milk of healthy primiparous women living in the Naples countryside was conducted. From 47 healthy primiparous women, breast milk samples were collected within the first six weeks after delivery as well as clinical data of mother–newborn pairs. Breast milk analyses were performed with both competitive indirect enzyme-linked immunosorbent assay and high-performance liquid chromatography with fluorescence detection. At 36.9 ± 2.6 days after vaginal delivery, the mean zearalenone contents of the breast milk samples were 1.13 ± 0.34 µg/L. The zearalenone levels correlated with both mother weights before pregnancy (r = ?0.506; P < 0.001) and at delivery (r = ?0.351; P < 0.05). The present results indicate that breast milk may be contaminated with zearalenone.     

Microbiological and mycotoxicological contamination in Slovenian primary grain production

This study was aimed at investigating the contamination of grains, which are grown and used for animal diets by Slovene farmers. A total of 107 samples were examined for mesofilic aerobic molds and yeasts. The leading contaminants among molds were from Fusarium spp., detected in 73 samples, mostly in maize (50). In accordance with the European Feed Microbiology Organisation criteria, 68% of all tested samples were of the quality level I, with normal microbiological flora, and 9% of samples were estimated as quality level IV and not suitable for animal feed. Using liquid and gas chromatography methods, the presence of various mycotoxins (expressed as 12% of moisture content) was found in 77% of all samples; mostly deoxynivalenol (71%) in concentration of 150–14,420 µg kg^?1, followed by zearalenone (32%, 20–640 µg kg^?1), fumonisin B1 (31%, 10–4863 µg kg^?1), fumonisin B2 (22%, 10–1629 µg kg^?1), while HT-2 and T-2 toxins were found in low concentration in two samples, and aflatoxin B1 and ochratoxin A in one sample. By statistical evaluation, significant positive correlations were noted between the presence of deoxynivalenol and zearalenone as well as between fumonisin B1 and B2.     

The role of tryptophan in don‐induced feed rejection

Abstract

Deoxynivalenol (DON, vomitoxin) commonly produced by Fusarium fungi can alter feeding behaviour of pigs and other farm animals. The effects of dietary tryptophan (TRP, precursor of brain amine serotonin) in combination with DON were examined in mice to determine if TRP can modulate DON toxicity. Results indicated that brain TRP can be influenced by dietary TRP, but no evidence of TRP potentiating DON toxicity was observed. Higher TRP levels likely induced amino acid imbalance leading to weight gain suppression.     

Effect of the appetite stimulant cyproheptadine on deoxynivalenol ‐ induced reductions in feed consumption and weight gain in the mouse 1

Abstract

Deoxynivalenol (DON, vomitoxin), a Fusarium mycotoxin, is suspected of inducing its anorectic/feed refusal activity through a serotoninergic (5HT) mechanism, possible via 5HT₂‐receptors. In this study the efficiency of cyproheptadine (CYP), a serotonin antagonist and known appetite stimulant, to attenuate the adverse effect of DON was investigated in mice. CYP was administered in the feed for two days before animals began receiving the DON, which was also added to the feed. Both agents were administered concurrently thereafter for a 12‐day period. Dosing levels included various combinations of the two compounds, ranging from 0–16 ppm DON and 0–20 ppm CYP.

Results showed that CYP could effectively offset the reduction in feed intake caused by dietary DON, but only when dose levels were optimized (CYP appeared to have a narrow effective dose range, which was also dependent on the DON concentration). In fact, optimum CYP doses were able to enhance feed intake in DON‐fed mice above control levels, indicating that more than just a direct block of the toxin's effect was occurring. Conversely though, the effect of CYP on weight gain (WG) was ambiguous. At the lower CYP doses (≤ 5 ppm), alone or in combination with the lowest DON level tested (4 ppm), a modest positive effect was noted, but at the higher DON concentrations (≥ 8 ppm) the overall WG generally remained depressed, and tended to be reduced further by increasing doses of CYP.

These results provide additional support that serotoninergic mechanisms probably play a role in mediating DON‐induced reduction in feed intake. Where and how DON actually initiates its anorectic effect, however, has not been resolved.     

Production of an antimicrobial cytochalasan by an endophytic Chaetomium globosum HYML55 from Hypericum mysorense and its RNA secondary structure analysis

The mutualistic benefits and disadvantages of non-clavicipitaceous endophytes for their hosts are not well studied and are postulated as being distinct from free-living or pathogenic counterparts. The defensive metabolites and antioxidants produced by endophytes are postulated to be the benefits offered for the hosts. Therefore, the antimicrobial and free-radical-scavenging activities of a foliar endophyte Chaetomium globosum HYML55 isolated from Hypericum mysorense of Western Ghats, India were studied. RNA secondary structure analysis refuted the unique biotope postulation because strain HYML55 was more similar to a coprophilous strain than to endophytic strains. Ethyl acetate extract of the fungus inhibited bacteria and fungi. The extract also quenched 98.65% 1,1-diphenyl- 2-picrylhydrazyl radicals with total antioxidant capacity of 13.62 mg ascorbic acid equivalent/g. A yellow amorphous antimicrobial compound purified from the extract was identified as chaetoglobosin F, which had a minimum inhibitory concentration of 7.8–15.6 μg/mL, except against Pseudomonas aeruginosa. This is the first report on the antimicrobial activity of chaetoglobosin F and other bioactivities of any endophytes from H. mysorense. The study emphasises the inter-habitat cycling of endophytes and possible benefits of C. globosum for the host. Chaetoglobosin F is an anti-inflammatory and anti-neoplastic compound, and C. globosum HYML55 could be exploited for the industrial production of this compound.     

A new hypothesis for the etiology of Kaschin-Beck disease

Kaschin-Beck disease (KBD) could be exogeneous free radical induced endemic osteoarthropathy under conditions of lacking necessary antioxidant defences.especially selenium-cored defence in Chinese eco-en-vironment. Experimental evidences have been provided for the internal relations among previously suggested pathologic factors, i.e., selenium deficiency, organic matters of drinking waterand mycotoxins from contaminated grains, based on the facts that the former is a preventory factor while the others are exogenous freeradical carriers. ESR spectrum shows that these exogenous free radical carriers have a property of semi-quinone compounds and are toxic in in-vitro cartilage cell culture and animal tests in inducing lipid peroxidation process.     

Cytotoxic and proinflammatory response of RAW 264.7 cells to differentially fractionated fungal fragments

Although significant research has been carried out for studying the impacts of pure mycotoxins on human cell, little information is known about the direct effects of fungal submicron particles. In this report, murine macrophage RAW 264.7 cells were exposed to fungal extracts from Stachybotrys chartarum RTI 5802 and Aspergillus versicolor RTI 3843 as well as mixtures from both fungi with fragment sizes ranging from 0.25 to 1?µm. Cell viability was found to be unaffected by all extracts except fragments in the 0.5–1?µm size range from A. versicolor RTI 3843, which decreased cell viability by approximately 60%. IL-1β expression was generally upregulated following exposure to fungal extracts from either S. chartarum RTI 5802 or A. versicolor RTI 3843. However, when extracts from the mixture of both fungi were introduced, IL-1β expression was downregulated. IL-6 and IL-10 expression were also generally upregulated. In comparison to the findings in the literature about the effects of pure mycotoxins on RAW 264.7 cells, the results presented herein show different patterns on mRNA expression which may be a result of the complex composition of the extract in terms of both mycotoxins and other fungal materials in addition to the potential interplay of the different components.