柠檬醛抑菌特性及其缓释制剂研究进展

随着环保理念的加强和可持续发展战略的实施推进,高效、低毒成为抑菌剂生产和使用的必然趋势。柠檬醛属于开链单萜类化合物,是山苍子、柠檬草等的次级代谢产物,对细菌、真菌、病毒等有很好的抑制作用,可以开发为天然植物源抑菌剂代替化学杀菌剂使用。然而柠檬醛具有不稳定,易挥发,光、热条件下易分解等缺点,这些缺点限制了柠檬醛的应用。缓释制剂能将被包埋的活性物质缓慢释放,同时对活性物质起到稳定和保护的作用。该文以柠檬醛为对象,围绕柠檬醛的抑菌活性、抑菌机理进行阐述,同时分析柠檬醛在应用上的不足,并对柠檬醛缓释制剂的种类和应用等进行归纳总结,为柠檬醛抑菌剂的开发和柠檬醛缓释制剂的应用提供参考。     

柠檬醛纳米胶囊的制备及其稳定性研究

以明胶、阿拉伯胶为壁材,司班60(S-60)、吐温20(T-20)为乳化剂,制备柠檬醛纳米胶囊。采用高压乳匀法,通过正交试验设计优化柠檬醛纳米胶囊的制备。采用动态光散粒径仪对其性能进行检测,利用顶空固相微萃取气质联用对柠檬醛主要降解产物进行研究。结果表明,壁材总量0.4%(明胶∶阿拉伯胶为1∶1),乳化剂总量0.4%(S-60∶T-20为2∶3),pH为3.8的缓冲液(1mol/L柠檬酸和1mol/L柠檬酸钠),柠檬醛0.1%,经120MPa压力均质3次。所制备得到的柠檬醛纳米胶囊体系稳定,平均粒径为228.4nm±7nm,柠檬醛纳米胶囊储藏6d(45℃)后柠檬醛剩余量为54.4%±0.2%。     

β-环糊精包埋柠檬醛微胶囊工艺优化及其缓释性能

以β-环糊精为壁材,采用乳化法制备柠檬醛微胶囊,考察固形物质量分数、乳化剂质量分数和心壁比对柠檬醛微胶囊包埋产率和包埋效率的影响。采用响应面法对3个单因素工艺参数进行优化,回归得到二次多项式模型极显著,模型的相关系数为0.966 1,优化试验工艺条件为固形物质量分数26%、乳化剂质量分数0.8%和心壁比1∶4。在此条件下进行验证实验,柠檬醛微胶囊包埋产率值为78.8%。缓释性能实验结果表明,微胶囊化可以明显降低柠檬醛的挥发速率,Avrami’s方程也可以对柠檬醛微胶囊释放过程进行较好地拟合。     

柠檬醛对副溶血性弧菌的抑制作用

有效预防和控制副溶血性弧菌对食品的污染对于保障公众健康具有重要意义。本研究首先利用琼脂稀释法测定7?种植物源活性物质（丁香酸、阿魏酸、绿原酸、硫辛酸、原儿茶酸、原儿茶醛和柠檬醛）对副溶血性弧菌的最小抑菌浓度,在此基础上选择柠檬醛进行后续实验;通过检测经柠檬醛处理后的副溶血性弧菌生长曲线、细胞膜电位、胞内ATP浓度、细胞膜完整性以及细胞形态的变化,探究柠檬醛对副溶血性弧菌的抑制作用及可能的作用机理。结果表明：柠檬醛相比于其他6?种植物源活性物质对副溶血性弧菌有更好的抑菌效果,其对两株标准菌株和8?株分离菌株的最小抑菌浓度在0.10～0.60?mg/mL范围内;柠檬醛能够引起副溶血性弧菌细胞膜电位去极化、胞内ATP浓度降低及细胞膜完整性下降,同时可使细胞皱缩变形。本研究结果表明柠檬醛具有良好的抑菌功效,并有潜力作为天然的抗菌物质应用于食品工业。     

β-环糊精稳定柠檬醛Pickering乳液的制备及其稳定性研究

制备由β-环糊精稳定的高比例柠檬醛相Pickering乳液并研究乳液的稳定特性。利用β-环糊精与柠檬醛的包合物微粒作为乳化稳定剂、通过高速均质法制备柠檬醛的Pickering乳液,观察乳液的微观结构、测定其粒度和Zeta电位,研究体系pH值及添加黄原胶对乳液稳定性的影响,并对其稳定性进行了初步测定。结果表明,柠檬醛-水体系经β-环糊精稳定乳化后可形成稳定的水包油型Pickering乳液,油滴呈标准球形,粒径在数百纳米;偏酸或偏碱性环境条件及添加黄原胶有利于乳液的稳定。进一步研究表明,乳液具有较好的耐热稳定性,但耐冻能力较差,黄原胶的加入(5 g/L)可显著提高乳液的耐冻性。该研究为柠檬醛乳液的绿色制备提供一种新的途径。     

喷雾干燥法制备柠檬醛微胶囊及微胶囊稳定性研究

以甲基纤维素（MC）、羧甲基壳聚糖（CMCTS）和海藻酸钠（Alg）为复合壁材,通过层层自组装法,采用喷雾干燥法制备柠檬醛微胶囊,研究了壁材组成对微胶囊的柠檬醛含量、形态、吸水性能及稳定性的影响,以柠檬醛含量为指标确定柠檬醛微胶囊的最佳壁材组成及工艺条件为:甲基纤维素（内层）0.8g/50mLH₂O、羧甲基壳聚糖1.0g/30mLH₂O、海藻酸钠6.0g/180mLH₂O、甲基纤维素（外层）0.8g/50mLH₂O,柠檬醛含量为4.54%。添加双亲性MC作为内层壁材使柠檬醛含量提高了3.12%。以MC作为外层壁材使柠檬醛含量提高了0.23%,90d内微胶囊的吸水量降低了0.42%,提高了微胶囊在贮存过程中的稳定性。      

β-环糊精包埋丁香酚、肉桂醛、柠檬醛缓释防腐包在传统肉干上的防霉效果对比

将具有抗菌活性的丁香酚、肉桂醛、柠檬醛用β-环糊精包埋后制成抗菌活性缓释防腐包,应用于传统肉干的保藏,通过观察肉干贮藏过程中霉变情况,研究其应用效果。本文根据肉干干燥过程特性,确定了肉干贮藏较适宜的水分含量为16.40%,水分活度为0.695,将此水分肉干作为样品,于28℃贮藏6个月后,未加入防腐包的空白对照组肉干28 d开始长出霉菌,而包合物添加量在2.0 g以上的丁香酚、肉桂醛、柠檬醛缓释防腐包的肉干组6个月均未长霉,具有较好的防霉变效果,但低于2.0 g防腐包添加量时,肉干贮藏一个月后,陆续长出霉菌,此时防腐效果为丁香酚>柠檬醛>肉桂醛。     

柠檬醛和辛醛混合物对指状青霉的抑制作用

本研究采用琼脂稀释培养法测定柠檬醛、辛醛以及柠檬醛+辛醛混合物(V/V,4:1)对指状青霉菌丝体生长的影响,并对柠檬醛+辛醛混合物抑菌机制进行初步探讨。结果表明,柠檬醛+辛醛混合物(V/V,4:1)能明显抑制指状青霉菌丝体生长,最小抑菌浓度(minimal inhibitory concentration,MIC)和最小杀菌浓度(minimal fungicidal concentration,MFC)均为2.50μL/m L,而柠檬醛的MIC和MFC分别为2.50μL/m L和5.00μL/m L,辛醛的MIC和MFC分别为0.63μL/m L和2.50μL/m L。扫描电镜结果显示,经MIC(MFC)的柠檬醛+辛醛混合物处理后,指状青霉菌丝体表面发生严重塌陷和多重褶皱,菌丝结构发生不规则扭曲,表明细胞完整性遭到严重破坏。此外,指状青霉菌丝体胞外p H、胞外电导率以及260 nm处的吸光度值均随柠檬醛+辛醛混合物(V/V,4:1)浓度增加显著增加。本研究结果表明,柠檬醛+辛醛混合物(V/V,4:1)能破坏指状青霉细胞结构的完整性,增加细胞膜通透性,导致细胞内物质泄漏,从而导致菌丝体死亡。     

茶树油、丁香酚和柠檬醛对荔枝黑曲霉的抑制作用

研究茶树油、丁香酚和柠檬醛的挥发性香氛对荔枝致腐真菌黑曲霉的抑制作用,采用复配方法探究3种香料对黑曲霉的协同抑菌性。结果表明：丁香酚和柠檬醛的挥发性香氛对黑曲霉有较强的抑制作用,最低抑菌浓度分别为0.3μL/cm3和0.4μL/cm3;茶树油挥发性香氛抑制黑曲霉的作用稍差,但与丁香酚和柠檬醛复配后协同抑菌作用增强。     

柠檬醛对花生酱的防腐效果

采用平板法比较柠檬醛与合成食用防腐剂苯甲酸钠、山梨酸钾对8种霉菌的抗菌效力。结果表明，在花生酱pH5.5时，柠檬醛对多数霉菌的最低抑菌浓度（MIC）为0．10％，山梨酸钾为0．10％，而苯甲酸钠的MIC值在0．35％或0．35％以上。同时，从柠檬醛与黄曲霉产毒关系的试验中发现，柠檬醛对黄曲霉产生黄曲霉毒素有较强的抑制作用。     

山苍子精油抑制黄曲霉菌的生长和产毒作用研究

山苍子精油是一种纯天然植物精油,本文研究了其对黄曲霉生长、代谢和毒素产生的抑制作用,探讨了山苍子精油对黄曲霉菌的抑菌能力和作用机理。本研究将花生放置于自然环境染菌并分离纯化目标菌,采用形态学并结合ITS序列法进行菌株分类鉴定;结合抑菌圈、抑菌率和最低抑菌浓度(MIC)的测定探讨山苍子精油对黄曲霉菌的抑制能力;进行了山苍子精油影响黄曲霉孢子萌发率、生长曲线和黄曲霉毒素B1产生的实验研究;从细胞膜渗透性、细胞酶活性的变化探讨了山苍子精油抑制黄曲霉的作用机理。实验结果表明:从腐败花生中分离筛选出菌株HB₂,经ITS序列法鉴定为黄曲霉(Aspergillus flavus);黄曲霉素测定结果显示其含有黄曲霉素B1(AFB1),质量浓度为3.4×10³μg·kg^-1(纯湿菌体);抑菌圈随精油浓度的增大明显变大,对黄曲霉的最低抑菌体积分数(MIC)为0.800μL·mL^-1;孢子萌发率、牙管长度、黄曲霉菌体的生长量和AFB1的浓度随培养液中精油浓度的增大呈显著下降趋势,当山苍子精油浓度为0.100μL·mL     

Impact of habitats on toxigenic potential of Aspergillus flavus

Toxigenic potentials of Aspergillus flavus strains isolated from different organic substrates, air and soils were evaluated. The frequency of occurrence of A. flavus ranged from 76% in maize to 87% (coconut and groundnut) and 92% in makhana (Euryale ferox indica.) Incidence was lowest in green gram (42%). Analysis of variance showed that the percentage incidence of A. flavus in the aerosphere of maize fields was significantly affected by season × location. In soil samples the frequency of occurrence of A. flavus was high during the monsoons (76% in non-diara land soils, 69% in diara land soil). Among 1706 isolates of A. flavus obtained from different sources, 826 (48.4%) were found to be toxigenic. The frequency of non-toxigenic strains of A. flavus was comparatively higher (ratio = 1.07:1) than the toxigenic strains. Percentage incidence of the toxigenic strains of A. flavus was the highest (73.3%; ratio = 0.36:1) in the soil samples. No attempt was made to differentiate A. flavus and A. parasiticus and therefore all references to A. flavus include A. parasiticus.     

Spectral analyses of fluorescences in dent maize infected with Aspergillus flavus and Aspergillus parasiticus

Bright greenish yellow (BGYF) and blue white (BWF) fluorescences were associated with Aspergillus flavus and A. parasiticus infected maize. The fluorescences were studied spectrofluorometrically, the BGYF exhibiting a peak wave length between 480–485 nm and the BWF between 440–445 nm. Neither fluorescence varied in maize stored under different moistures and temperatures.

BWF was similar spectrally to the fluorescence of the endosperm of sound kernels but × 5 20 more intense. The spectrum of BWF was similar to Aflatoxin G₁ or a mixture of aflatoxin B₁, B₂, G₁ and G₂ when they were spotted on endosperm tissue. A color reference for BGYF was similar in peak wave length to BGYF. Amsoy soybeans without the seed coat fluoresced with a peak 470–475 nm and the intensity was low compared to BGYF in maize. A fluorescence of maize kernels visually similar to BGYF but not associated with Aspergillus infection or aflatoxin contamination was also investigated. This “false BGY” fluorescence was spectrally similar to the BGYF in infected kernels.     

Aflatoxin production by Aspergillus flavus in Brazil nuts

Experiments were conducted to evaluate the effects of relative humidity (r.h.; 75%, 80%, 85%, 97%) and temperature (10, 13, 15, 25, 30 °C) on aflatoxin production in previously dried (3.5% moisture content; m.c.) Brazil nuts. Initially Aspergillus spp. were isolated from the surfaces of whole in-shell (WIS) Brazil nuts imported from Peru using A. flavus and A. parasiticus agar (AFPA). Isolates were subsequently screened for aflatoxin production using yeast extract sucrose medium. Total aflatoxin (B₁+B₂+G₁+G₂+M₁) was analyzed using an immunoassay technique while the presence of aflatoxin was confirmed using thin-layer chromatography. The surface of shelled half-nuts (simulating damaged or trimmed nuts), shelled whole (SW) nuts, and WIS nuts following a chlorine wash and water rinse, served as sites for inoculation (10 μl; 10⁵/ml) using an aflatoxigenic isolate. Maximum concentrations of total aflatoxin and B₁ were detected in nuts stored at 97% r.h. and at temperatures of 25–30 °C. Shelled half-nuts contained the highest total (6817 ng/g) and B₁ (4483 ng/g) aflatoxin. WIS nuts contained the least total and B₁ toxin with maximum concentrations of 93 and 49 ng/g, respectively. Aflatoxin was not detected (detection limit of 1.75 ng/g) in nuts maintained at either 10 °C (97% r.h.) or at 30 °C (75% r.h.) for up to 60 d. Maximal moisture contents (%) and water activity values (a_W) for nuts stored at these conditions were 4.50 and 0.78, and 9.14 and 0.92, respectively. Results of this study indicate that the limiting moisture content and a_W values required to control aflatoxin production () in SW and WIS stored at 30 °C for up to 60 d are 4.5, 0.68, 5.0, and 0.75, respectively. Overall, increasing the relative humidity and temperature during storage resulted in an increase in aflatoxin and these were shown to be the most significant variables influencing toxin production in Brazil nuts.     

钾离子通道在黄曲霉菌孢子萌发中的作用

黄曲霉毒素严重污染农产品,威胁人畜生命健康,而孢子萌发是黄曲霉菌生存及侵染过程中的决定性事件,从最上游阻断孢子萌发可能是有效控制黄曲霉污染、开发新型抗菌药物的突破口之一。研究发现一定浓度的胞外KCl能够影响黄曲霉孢子的萌发。通过使用K+特异性荧光探针PBFI-AM,发现适合孢子萌发的特定浓度K+能够激发黄曲霉孢子瞬间产生较高的钾离子流,说明K+响应可能是黄曲霉孢子萌发的早期信号。4-AP作为特异性电压门控钾离子通道阻滞剂,能够明显抑制并延迟孢子的萌发。通过生物信息学预测,发现黄曲霉菌KCNA具有钾离子通道选择器序列,具备电压门控钾离子通道的α亚基属性。进一步构建KCNA非洲爪蟾卵母细胞表达体系,用双电极电压钳系统测得KCNA外向型电流,一定程度上证明了KCNA电压门控钾离子通道蛋白质属性,也说明电压门控钾离子通道参与了黄曲霉孢子萌发。     

非脱羧勒克菌wt16对黄曲霉菌生长与产毒的抑制作用

黄曲霉菌及其毒素严重威胁农产品质量安全,本文旨在探寻非脱羧勒克菌对黄曲霉菌及其毒素污染防控效果。从湖北黄陂分离筛选出一株非脱梭勒克菌wt16,将其与黄曲霉菌在液体培养基中共培养后测定非脱羧勒克菌wt16菌株对黄曲霉菌生长及产毒的抑制率。结果表明,在沙氏液体培养基中,非脱羧勒克菌wt16能明显抑制黄曲霉菌的生长及产毒,对其菌丝生长的抑制率为77%~92%,对其产毒的抑制率为90%~96%。通过扫描电子显微镜观察发现,非脱羧勒克菌wt16能改变黄曲霉菌丝的形态,使得黄曲霉菌丝体由规则的球体聚集成不规则形状,单个菌丝会由细长型断裂成小截形态,菌丝表面也变得更为光滑;并且发现wt16在花生粉及未受机械损伤的花生颗粒上对黄曲霉菌的生长及产毒均表现出很强的抑制作用。进一步研究发现,非脱羧勒克菌wt16菌株发酵上清液中含有能抑制黄曲霉毒素合成的有效成分,且该发酵上清液的制备以培养4 d以上为最佳,培养温度为15~40 ℃。     

Mature seeds of Sorghum contain proteins toxic to aflatoxin-producing Aspergillus flavus

Crude acid extract of mature Sorghum seeds showed partial inhibition of spore germination and growth of aflatoxin-producing Aspergillus flavus strains. To find the protein components involved in this mechanism of inhibition, the crude extract was fractionated by differential precipitation followed by ion exchange chromatography in carboxymethyl cellulose. Five proteins of low molecular weight were isolated. The ability of each of the five proteins to inhibit the spore germination and hyphal extension of aflatoxin-producing Aspergillus flavus strains was tested. Protein 5 did not show any inhibition. Proteins 1, 2 and 4 showed complete inhibition of spore germination (1500–2000 spores/100 μl of medium) at a concentration of 15 μg/100 μl. Protein 3 showed partial inhibition only.     

植物精油对黄曲霉的抑制机理及应用方式研究进展

黄曲霉是一种常见的真菌,其产生的黄曲霉毒素会对人和动物的生命健康造成威胁,近年来黄曲霉毒素中毒事件频繁发生。植物精油作为一种天然的活性物质,对黄曲霉有较强的抑制作用,其抑菌机理和应用方式依旧受到各国科研工作者广泛关注。本文主要概述了黄曲霉的危害,植物精油对黄曲霉的抑制效果及作用机理,总结了近几年植物精油抑制黄曲霉的应用方式,以期为植物精油在抑制黄曲霉中的合理应用提供理论依据。     

Aspergillus flavus link and its occurrence in relation to other mycoflora on stored spices

Spices are used sparingly to flavour foods; their mycoflora and its possible role in food infection has not been thoroughly investigated. In an examination of 6 spices, Aspergillus flavus, other Aspergillus species, and species of Rhizopus, Mucor, Penicillium, Tricothecium, and Fusarium were found on seed surfaces. A. flavus which is implicated in the production of carcinogenic aflatoxin, was mainly found on ginger, mustard, garlic, and pepper. The highest fungal counts (10.6 × 10⁴/g) occurred in stored pepper (Piper nigrum L.) and the lowest in curry leaves (Murraya koenigii L.). Contamination of pepper and mustard with A. flavus seems to be related to the processing and storage of these spices.