The impact of roasting on the ochratoxin A content of coffee

Roasting coffee led to a drop in the ochratoxin A (OTA) concentration, as measured by the reference method, especially for dark type roasts. The way the beverage was prepared also affected the OTA content, which could paradoxically be higher than that of the initial roasted coffee. Assays on the thermal stability of pure OTA showed that it ought to be found in larger quantities in roasted coffee. This suggested that OTA was masked by reactions with the substrate during roasting. The absence of OTA in green coffee is therefore the best guarantee of safety.     

Ecophysiology of ochratoxigenic Aspergillus ochraceus and Penicillium verrucosum isolates. Predictive models for fungal spoilage prevention - a review

Ochratoxin A (OTA) is a secondary metabolite produced by several species of Aspergillus and Penicillium; among them Aspergillus ochraceus and Penicillium verrucosum are two ochratoxigenic species capable of growing in different climates and thus contamination of food crops with OTA can occur worldwide. OTA can be found in a wide range of foods such as cereals, coffee, cocoa, spices, beer, wine, dried vine fruit, grapes and meat products. OTA is toxic to animals, it presents neurotoxic, immunotoxic and nephrotoxic effects. It has been implicated in a human kidney disorder known as Balkan Endemic Nephropathy. This review focuses on the ecophysiology of ochratoxin-producing Aspergillus ochraceus and Penicillium verrucosum, the effect of environmental factors on their germination, mycelial growth, and OTA production. Knowledge of environmental conditions required for sucessive stages of fungal development represent the first step towards preventing mycotoxin formation. Predictive models for different stages of fungal development are presented, which allow prediction of the time before spoilage as a function of the abiotic factors. Finally, the implications of these studies in management of barley, coffee and grapes are described. This can help to identify the critical control points in their production, storage and distribution processes.     

Banana peel: an effective biosorbent for aflatoxins

This work reports the application of banana peel as a novel bioadsorbent for in vitro removal of five mycotoxins (aflatoxins (AFB1, AFB2, AFG1, AFG2) and ochratoxin A). The effect of operational parameters including initial pH, adsorbent dose, contact time and temperature were studied in batch adsorption experiments. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and point of zero charge (pH_pzc) analysis were used to characterise the adsorbent material. Aflatoxins’ adsorption equilibrium was achieved in 15 min, with highest adsorption at alkaline pH (6–8), while ochratoxin has not shown any significant adsorption due to surface charge repulsion. The experimental equilibrium data were tested by Langmuir, Freundlich and Hill isotherms. The Langmuir isotherm was found to be the best fitted model for aflatoxins, and the maximum monolayer coverage (Q₀) was determined to be 8.4, 9.5, 0.4 and 1.1 ng mg^?1 for AFB1, AFB2, AFG1 and AFG2 respectively. Thermodynamic parameters including changes in free energy (ΔG), enthalpy (ΔH) and entropy (ΔS) were determined for the four aflatoxins. Free energy change and enthalpy change demonstrated that the adsorption process was exothermic and spontaneous. Adsorption and desorption study at different pH further demonstrated that the sorption of toxins was strong enough to sustain pH changes that would be experienced in the gastrointestinal tract. This study suggests that biosorption of aflatoxins by dried banana peel may be an effective low-cost decontamination method for incorporation in animal feed diets.     

Fermentation of table cream by Lactobacillus plantarum strains: effect on fungal growth,aflatoxin M1 and ochratoxin A

Aflatoxin M1 (AFM₁) and ochratoxin A (OTA) are two main mycotoxins in milk and dairy products. In the present work, the ability of four Lactobacillus strains (L. plantarum PTCC 1058, L. plantarum LP3, L. plantarum AF1 and L. plantarum LU5) to remove AFM1 and OTA in fermented cream was studied during 24 h fermentation. The antifungal activity of the mentioned lactobacilli against the defined fungi (Aspergillus flavus PTCC 5004, Aspergillus parasiticus PTCC 5018, Aspergillus nidulans PTCC 5014, Aspergillus ochraceus PTCC 5060) was also evaluated. The results showed that the cell counts of all strains were increased by 64–70% during fermentation. All Lactobacillus strains decreased the amount of AFM₁ significantly (P ≤ 0.05) in the range of 26–52%, which the highest AFM₁-reducing effect was related to L. plantarum LU5 (from 0.5 to 0.24 μg kg⁻¹). The mean OTA removal by Lactobacillus strains in fermented cream also ranged from 32 to 58%. Amongst Lactobacillus strains, the cell-free culture supernatants of L. plantarum LU5 showed the highest (inhibition zone of 26.7 ± 1.2 mm) and L. plantarum LP3 and L. plantarum PTCC 1058 the lowest antifungal activities. The fermented creams contained Lactobacillus strains exhibited the highest and lowest antifungal activities against A. ochraceus and A. parasiticus, respectively. L. plantarum LU5, with the inhibition zone of 27.6 ± 0.9 mm, was the most effective fungal inhibitor, while L. plantarum PTCC 1058 had the lowest antifungal activity.     

Effects of feeding bentonite clay upon ochratoxin A–induced immunosuppression in broiler chicks

A presence of mycotoxins in feed is one of the most alarming issues in the poultry feed industry. Ochratoxins, produced by several Aspergillus and Penicillium species, are important mycotoxin regarding the health status of poultry birds. Ochratoxins are further classified into to several subtypes (A, B, C, etc) depending on their chemical structures, but ochratoxin A (OTA) is considered the most important and toxic. Bentonite clay, belonging to phyllosilicates and formed from weathering of volcanic ashes, has adsorbent ability for several mycotoxins. The present study was designed to study the effects of bentonite clay upon OTA-induced immunosuppression in broiler chicks. For this, 480 day-old broiler chicks were procured from a local hatchery and then different combinations of OTA (0.15, 0.3, or 1.0 mg/kg) and bentonite clay (5, 10, and 20 g/kg) were incorporated into their feed. At 13, 30, and 42 days of age, parameters such as antibody responses to sheep red blood cells, in situ lymphoproliferative responses to mitogen (PHA-P), and in situ phagocytic activity (i.e., via carbon clearance) were determined respectively. The results indicated there was a significant reduction of total antibody and immunoglobulin titres, lymphoproliferative responses, and phagocytic potential in OTA-treated birds, suggesting clear immunosuppression by OTA in birds in a dose-dependent manner. These results were also significantly lower in all combination groups (OTA with bentonite clay), suggesting few to no effects of feeding bentonite clay upon OTA- induced alterations in different immune parameters.     

赭曲霉毒素A单克隆抗体的体外制备及其重组抗体的构建研究

目的 实现抗赭曲霉毒素A(ochratoxinA,OTA)单克隆抗体的体外制备及其重组抗体表达载体的构建与瞬时转染表达。方法 以前期获得的OTA杂交瘤细胞株(O16)为研究对象,通过无血清驯化培养的方式开展OTA单克隆抗体的体外制备研究;采用简并引物法获取OTA杂交瘤细胞的单克隆抗体编码基因,在此基础上构建OTA重组全长抗体表达载体以及OTA重、轻链可变区基因片段与人源恒定区片段连接的重组嵌合抗体表达载体,并基于哺乳动物细胞系,开展两种重组抗体的瞬时转染表达研究。结果 通过缓降血清浓度的方式对OTA杂交瘤细胞进行无血清驯化,实现了体外培养制备OTA单克隆抗体;在此基础上测得OTA单克隆抗体的重链和轻链基因序列,并成功构建了OTA重组全长抗体表达载体(pCDNA3.4-OTA-H/L)和重组嵌合抗体表达载体(p FUSE-OTA-H/L),通过共转染实现了两者在哺乳动物细胞系中的瞬时转染表达;经间接酶联免疫吸附测定(enzyme linked immunosorbent assay, ELISA)确定体外培养制备的OTA抗体、OTA重组全长抗体和嵌合抗体均具备特异性结合抗原的能力,三者的...     

同时检测玉米中黄曲霉毒素B1和赭曲霉毒素A的时间分辨荧光免疫层析试纸条的研制

研究制备了一种可同时快速定量检测黄曲霉毒素B1 (aflatoxin B1,AFB1)和赭曲霉毒素A(ochratoxin A,OTA)的二联时间分辨荧光免疫层析试纸条.采用铕系时间分辨荧光微球分别标记AFB1和OTA单克隆抗体,优化荧光微球活化pH值、标记的抗体浓度、荧光探针使用量、检测线包被原浓度、质控线羊抗鼠Ig...     

A Review on Mycotoxins in Food and Feed: Malaysia Case Study

Fungi are distributed worldwide and can be found in various foods and feedstuffs from almost every part of the world. Mycotoxins are secondary metabolites produced by some fungal species and may impose food safety risks to human health. Among all mycotoxins, aflatoxins (AFs), ochratoxin A (OTA), trichothecenes, deoxynivalenol (DON and T‐2 toxin), zearalenone (ZEN), and fumonisins (FMN) have received much attention due to high frequency and severe health effects in humans and animals. Malaysia has heavy rainfall throughout the year, high temperatures (28 to 31 °C), and high relative humidity (70% to 80% during wet seasons). Stored crops under such conditions can easily be contaminated by mycotoxin‐producing fungi. The most important mycotoxins in Malaysian foods are AFs, OTA, DON, ZEN, and FMN that can be found in peanuts, cereal grains, cocoa beans, and spices. AFs have been reported to occur in several cereal grains, feeds, nuts, and nut products consumed in Malaysia. Spices, oilseeds, milk, eggs, and herbal medicines have been reported to be contaminated with AFs (lower than the Malaysian acceptable level of 35 ng/g for total AFs). OTA, a possible human carcinogen, was reported in cereal grains, nuts, and spices in Malaysian market. ZEN was detected in Malaysian rice, oat, barley, maize meal, and wheat at different levels. DON contamination, although at low levels, was reported in rice, maize, barley, oat, wheat, and wheat‐based products in Malaysia. FMN was reported in feed and some cereal grains consumed in Malaysia. Since some food commodities are more susceptible than others to fungal growth and mycotoxin contamination, more stringent prevention and control methods are required.     

酶联适体分析法检测葡萄酒中的赭曲霉素A

建立了竞争取代酶联适体分析方法,检测葡萄酒中的赭曲霉素A(OTA)。核酸适体和OTA特异性结合导致与核酸适体杂交的短链DNA解链,解离的DNA作为捕获元素,进一步特异性结合辣根过氧化物酶(HRP),HRP催化四甲基二苯胺(TMB)底物显色,测定A450nm与浓度的线性关系,确定OTA的检出限。考察了DNA包被浓度、杂交温度和封闭液等因素对检测的影响。结果表明,在优化的条件下,所建立的竞争取代酶联适体分析法对OTA检测有高灵敏度,检测限0.88μg/L,线性范围1~100μg/L在葡萄酒中添加时,加标回收率为92.03%~106.6%,相对标准偏差RSD(n=5)小于2.1%,此方法可用于葡萄酒中OTA的快速测定。     

赭曲霉毒素A生成转化及致毒机制的研究进展

赭曲霉毒素A(Ochratoxin A,OTA)是由曲霉属(Aspergillus.sp)和青霉属(Penicillium.sp)真菌产生的一种次级代谢产物,它的生成受温度、水活度等的影响。检测食品及饲料中OTA含量的基本方法有薄层层析法、高效液相色谱法和酶联免疫吸附法。OTA因被认为与巴尔干半岛肾病有关而引起全球的关注,研究发现,OTA具有肾毒性、肝毒性、免疫毒性、基因毒性等,并且主要是通过促进膜的过氧化反应,抑制线粒体的呼吸作用和影响细胞信号传导通路中蛋白及关键因子的转录表达等来达到致毒效应。吸附、转化、降解是OTA脱毒的主要方式。本文就OTA的检测方法、生物合成、致毒机制和脱毒转化的相关研究进展进行了综述。