欧姆加热对嗜酸耐热菌的电穿孔效应

探讨了欧姆加热对嗜酸耐热菌的杀灭作用机理.用自行设计的批式欧姆加热装置,对苹果汁中的嗜酸耐热菌进行处理后,利用扫描电镜观察其细胞壁膜结构的变化,利用电导率仪和紫外吸收分光光度计测定菌悬液的成分变化.结果表明:通过环境扫描电镜可观察到嗜酸耐热菌的表面出现凹陷和破损,电导率仪和紫外吸收分光光度计测定细胞内容物的溢出,可以推断出欧姆加热造成了嗜酸耐热菌的"电穿孔".     

苹果法的欧姆加热研究

研究了苹果汁的欧姆加热规律及其对果汁品质的影响,考察了温度(T)和可溶性固形物(SS)含量对电导率σ的影响,以及电场强度(E)、可溶性固形物(SS)和加热体积(液高 h)对加热速率的影响、同时考察了欧姆加热对果汁糖度、pH、透光率等的影响.结果表明,苹果汁的电导率随温度的升高而增大,且呈线性关系,随着可溶性固形物含量的增加,苹果汁的电导率和加热速率都是先增大然后减小,在26°Bx左右时最大.苹果汁的加热速率随电场强度的增加而增大,而在相同电场强度下,液高对加热速率没有影响.并且,欧姆加热对果汁品质影响很小,因此极具开发前景.     

藏花素理化性能的测试

藏花素是从茜草科植物栀子的果实中提取精制的一种天然食用黄色素。本文对藏花素的理化性能作了全面的测试,用紫外一可见分光光度计,红外光谱仪测定了该色素的吸收光谱。用紫外一可见分光光度计测定pH值、金属离子、热、光等因素对藏花素的影响。用原子吸收分光光度计测定了藏花素中As、Pb含量。从而得出:藏花素溶解性好,着色力强、金属离子、酸、碱、糖对色调无影响;耐热、耐光、As、Pb含量很低,是一种比较理想的天然食用黄色素。     

苹果汁电导率及介电损耗因子影响因素的研究

本文研究了苹果汁的电导率与可溶性固形物含量、温度、粘度和NaCl浓度等因素的关系,并通过电导率与介电损耗因子的关系,确定不同频率下与离子介导相关的苹果汁的介电损耗因子。结果表明,苹果汁的电导率随可溶性固形物含量的增加呈先增大后减小的抛物线趋势,随温度线性增大,粘度大小对电导率的影响很小,添加少量NaCl苹果汁电导率成倍增大。借助SPASS数据分析软件,得出电导率同可溶性固形物含量、温度的非线性回归方程：σ=-28.69C2+17.292C+0.02T+0.135,R2为0.974,拟合度良好。通过计算获得苹果汁在不同频率下（13.56、27.12、40.68 MHz）与离子介导相关的介电损耗因子。这些结果和数据,可为欧姆和射频加热技术用于苹果汁生产提供基础数据支持。     

大豆酱欧姆加热影响因素的初步研究

研究了不同欧姆加热条件对大豆酱电导率、加热速率以及酵母菌杀菌效果的影响.实验结果表明大豆酱的加热速率随着电压、盐含量的增大和固形物含量降低而增大,大豆酱的电导率与温度呈线性关系;正交试验确定最佳杀菌条件65℃、150s、35V;在此条件下,能全部杀灭大豆酱中的酵母菌,电能消耗功率约为785J/kg·s.     

柠檬酸及其盐类中嗜酸耐热菌的检测

目的对柠檬酸及其盐类中嗜酸耐热菌进行培养,研究最佳检测方法。方法通过改变培养基种类、pH值和热休克方式三个因素,以菌落数多少进行综合评价,优选最佳检测方法。结果检测嗜酸耐热菌的最佳条件:70℃热处理20 min,室温水冷却,用滤膜抽滤后接种于pH值为3.5~4.0的YSG培养基上,置45℃培养5 d。结论该方法对柠檬酸及其盐类中嗜酸耐热菌的检测有较强的实用性。     

气相色谱-质谱法分析乳酸菌发酵苹果汁香气成分

以乳酸菌单一菌种和多菌组合分别发酵苹果汁,采用紫外-可见分光光度计测定发酵过程中苹果汁光密度(optical density,OD)的变化,采用顶空固相微萃取和气相色谱-质谱联用技术检测9组发酵苹果汁的香气成分,结合不同香气物质的阈值与香气值分析特征香气成分,并对各组苹果汁进行感官评价。结果表明:在发酵过程中苹果汁的OD值呈对数上升趋势。各组苹果汁中共鉴定出48种香气成分,主要包含醇类、酯类、醛类、酮类和酚类,其中酯类是各发酵组中种类最多且含量最大的香气类别,其次是醇类。9组苹果汁的主要香气成分和香气值较大并有突出贡献的特征香气成分均为丁酸乙酯、2-甲基丁酸乙酯、2-甲基丁基乙酸酯和乙酸己酯等。多菌发酵和单菌发酵苹果汁的香气成分含量及香气值差异显著(P0.05),说明多菌发酵中醇类、酯类和其他类香气物质的总含量明显高于单菌发酵,且赋予苹果汁更强烈的果香、青香和花香。多菌发酵组的总体感官评分高于单菌发酵组,当副干酪乳杆菌20241、动物双歧杆菌6165、嗜热链球菌6063、嗜酸乳杆菌6005活菌数比为1∶1∶1∶1时,感官评分最优。研究结果为今后乳酸菌在发酵果蔬汁方面的研究和应用提供理论依据。     

欧姆加热的电参数对杀菌效果的影响

采用欧姆加热杀菌与水浴加热杀菌对比的方法。以豆浆的总菌数为试验菌群,通过改变电源的电压和频率,得出电源的电压和频率与杀菌效果的关系。对比水浴加热杀菌的效果可知:当电压超过60V时,欧姆加热方法的杀菌效果明显好于水浴加热方法的杀菌效果,残菌率随处理电压的增加以指数规律减少;随着频率的增加,欧姆加热的杀菌好,但当频率超过一定值时杀菌效果降低;频率在30～300Hz时其杀菌效果明显优于水浴加热方法的杀菌。     

海狗油成分及结构分析

利用原子吸收分光光度计和高效液相色谱仪分别对海狗油中的金属元素和维生素进行分析和鉴定,并借助紫外分光光度计、红外光谱仪和气相色谱-质谱连用仪对其脂肪酸结构、组成和含量进行初步确证.     

电导率法淀粉糊化温度测定及其影响因素的研究

改进通电加热淀粉悬浮液过程中电导率的测定装置,有效地防止淀粉悬浮液的沉淀及水分散失,提高测量精度,使之可用于淀粉糊化温度的测定,并将结果与差示扫描热量仪fDSCl测量结果进行了比较.并进一步研究淀粉浓度、NaCl浓度和电压对淀粉悬浮液的电导率、加热速率及所测得糊化温度的影响.结果表明:通电加热淀粉悬浮液时,随NaCl浓度的增加,所测得的糊化温度逐渐增大.随淀粉浓度的升高,所测得的糊化温度逐渐降低.随着电压的增大,淀粉悬浮液的电导率和加热速率也随之增大.改进后的装置可准确测定淀粉的糊化温度.与DSC结果相近.     

溶菌酶协同高压CO_2对酸土脂环酸杆菌芽孢的致死效果

以芽孢致死率为评价指标,通过单因素和正交试验考察溶菌酶添加量、压力、温度和时间在高压CO2处理过程中对酸土脂环酸杆菌芽孢致死效果的影响,并优化杀菌条件。结果显示:溶菌酶添加量、压力和温度对芽孢致死效果的影响均显著(P0.05),在溶菌酶添加量0.10g/kg、压力25MPa、温度55℃、时间60min的优化条件下芽孢致死率可达到5.47个数量级。另对此条件下处理后的样品进行高效液相色谱(HPLC)和扫描电子显微镜(SEM)检测,结果显示:芽孢核心物质2,6-吡啶二羧酸(DPA)发生大量泄露,且部分芽孢完全破裂,溶菌酶对高压CO2杀灭芽孢的协同效果显著(P0.05)。     

介质阻挡放电等离子体对不同pH值生长的酸土脂环酸芽孢杆菌的灭活作用

该研究探讨了介质阻挡放电等离子体对不同pH值（3.2、3.6、4.0和4.4）下生长的酸土脂环酸芽孢杆菌的杀灭作用。研究发现等离子体对最适生长pH值4.0下的酸土脂环酸芽孢杆菌杀灭效率最低,pH值升高（4.4）或降低（3.2、3.6）均引起致死率增加（P<0.05）。例如30 kV处理2 min条件下,等离子体对pH值为4.0生长的酸土脂环酸芽孢杆菌致死率为4.8个对数,对pH值为3.2、3.6和4.4生长的菌体致死率分别为5.9、5.4和5.1个对数。等离子体对不同pH值培养的酸土脂环酸芽孢杆菌致死率的差异可能与菌体形态和膜脂质成分有关。扫描电镜观察发现,pH值3.2和3.6培养的菌体出现异常伸长形态,且酸土脂环酸芽孢杆菌在pH值3.2条件下更为明显,更易被等离子损伤而导致死亡。气相-质谱联用结果表明相比于最适pH值4.0,pH值4.4生长条件下环己烷十一酸相对含量显著提高（67.86%）,而环戊烷十三酸降低（22.21%）（P<0.05）,使得等离子体产生的氧自由基等成分在细胞膜中的堆积,而导致菌体更易被杀灭。该研究表明菌体耐受性增加与酸土脂环酸芽孢杆菌细胞膜脂肪酸成分和形态变化存在一定的相关性。     

短波紫外发光二极管处理对脂环酸芽孢杆菌的灭活效果及作用机制

短波紫外发光二极管（ultraviolet-C light-emitting diode，UVC-LED）处理是一种新型的非热杀菌技术。本实验以果汁中常见的致腐菌脂环酸芽孢杆菌（Alicyclobacillus acidoterrestris）为目标菌，研究UVC-LED对脂环酸芽孢杆菌的杀灭作用，通过测定处理后细菌胞内核酸和蛋白质泄漏量、细胞膜通透性、胞内活性氧（reactive oxygen species，ROS）的累积水平以及胞内蛋白质和DNA的损伤情况，进一步探究UVC-LED对脂环酸芽孢杆菌的杀菌机理。结果表明：增加UVC-LED的照射剂量可增强其对脂环酸芽孢杆菌的杀灭效果，当照射剂量增加至50 mJ/cm2时，生理盐水中存活的细菌数量降低4.6（lg（CFU/mL））。通过对存活曲线的模拟，发现UVC-LED对生理盐水中脂环酸芽孢杆菌的杀灭作用既符合log-linear模型，又符合Weibull模型。处于不同生长时期的细菌对UVC-LED的敏感度不同，其中处于对数期的细菌对UVC-LED更敏感。照射处理导致膜通透性的改变以及内容物的泄漏，说明细胞膜结构遭到一定程度的破坏，但是胞内ROS的累积水平没有显著提高（P＞0.05），拉曼光谱分析表明胞内蛋白结构有所改变，经吖啶橙（acridine orange，AO）染色荧光显微镜观察发现照射处理后菌体DNA的结构变化明显。综上，UVC-LED可通过造成DNA损伤、蛋白结构变化和细胞膜透性改变从而杀灭脂环酸芽孢杆菌，根据破坏程度的不同，推测DNA损伤是细胞死亡的主要原因。     

分光光度计法快速检测果汁中嗜热耐酸菌

酸土脂环酸芽孢杆菌(Alicyclobacillus acidoterrestris)是果汁污染的主要嗜酸耐热菌。它可以将香草酸或者香兰素转化为愈创木酚,后者在过氧化物酶的作用下产生红褐色产物,从而可以快速检测该菌。根据这个原理,设计实验探究该颜色反应的最优培养条件以及实际体系中检测该菌的可行性。结果表明：在静止培养情况下,在BAM培养基中添加100μg/kg的香草酸最有利于愈创木酚的产生;不同浓度的嗜热耐酸菌芽孢液在上述条件下培养,随着浓度的降低产生高浓度愈创木酚时间依次延迟,最低浓度为3个/mL的芽孢培养21h后,愈创木酚含量用分光光度计测定的光密度值OD470为1.0以上,为肉眼可见的红褐色;模拟实际体系下的培养也出现了颜色反应,证明该方法确实可以实现果汁中嗜热耐酸菌的快速、简便、高灵敏度的检测。     

Guaiacol production from ferulic acid, vanillin and vanillic acid by Alicyclobacillus acidoterrestris

Alicyclobacilli are thermophilic, acidophilic bacteria (TAB) that spoil fruit juice products by producing guaiacol. It is currently believed that guaiacol is formed by Alicyclobacillus in fruit juices as a product of ferulic acid metabolism. The aim of this study was to identify the precursors that can be metabolised by Alicyclobacillus acidoterrestris to produce guaiacol and to evaluate the pathway of guaiacol production. A. acidoterrestris FB2 was incubated at 45°C for 7days in Bacillus acidoterrestris (BAT) broth supplemented with ferulic acid, vanillin or vanillic acid, respectively. The samples were analysed every day to determine the cell concentration, the supplement concentration using high performance liquid chromatography with UV-diode array detection (HPLC-DAD) and the guaiacol concentration, using both the peroxidase enzyme colourimetric assay (PECA) and HPLC-DAD. The cell concentration of A. acidoterrestris FB2 during the 7days in all samples were above the critical cell concentration of 10(5)cfu/mL reportedly required for guaiacol production. The guaiacol produced by A. acidoterrestris FB2 increased with an increase in vanillin or vanillic acid concentration and a metabolic pathway of A. acidoterrestris FB2 directly from vanillin to guaiacol was established. The high concentration of vanillic acid (1000mg/L) resulted in an initial inhibitory effect on the cells, but the cell concentration increased after day 2. Guaiacol production did not occur in the absence of either a precursor or A. acidoterrestris FB2 and guaiacol was not produced by A. acidoterrestris FB2 in the samples supplemented with ferulic acid. The presence of Alicyclobacillus spp. that has the ability to produce guaiacol, as well as the substrates vanillin or vanillic acid is prerequisite for production of guaiacol.     

Characterization of a wild strain of Alicyclobacillus acidoterrestris: heat resistance and implications for tomato juice

This article reports the characterization of a wild strain of Alicyclobacillus acidoterrestris and describes the implications of the heat resistance of this microorganism in tomato juice. The strain (labeled as A. acidoterrestrisγ4) showed pH and temperature ranges for growth typical of the species (3.0 to 6.0 for the pH and 35 to 60 °C for the temperature); heat resistance in tomato juice was as follows: D(T) values of 40.65, 9.47, and 1.5 min (at 85, 90, and 95 °C, respectively) and z-value of 7 °C. A treatment at 70 °C for 15 min was found to be optimal for spore activation, whereas Malt Extract Agar, acidified to pH 4.5, showed good results for spore recovery. Concerning the implications of heat resistance of A. acidoterrestris on tomato juice, high temperatures required for spore inactivation determined a general decrease of the antioxidant activity (increase of the redox potential and reduction of the chain-breaking activity), but not the formation of brown compounds (namely, hydroxymethylfurfural), thus suggesting an effect on the secondary antioxidants (carotenoids and ascorbic acid) rather than on lycopene. PRACTICAL APPLICATION: Alicyclobacillus acidoterrestris is an emerging spore-forming microorganism, capable of causing spoilage in tomato juice. Due to their high thermal resistance, spores could be used as targets for the optimization of heat processing; this article reports on the assessment of thermal resistance of a wild strain of A. acidoterrestris, then focusing on the effect of the thermal treatment necessary to inactivate spores on the quality of tomato juice.     

Thermal inactivation of Alicyclobacillus acidoterrestris spores under different temperature, soluble solids and pH conditions for the design of fruit processes

Alicyclobacillus acidoterrestris, a thermoacidophilic, non-pathogenic and spore-forming bacterium has been detected in several spoiled commercial pasteurised fruit juices. A. acidoterrestris spores, besides being resistant to the pasteurisation treatment conditions normally applied to acidic fruit products, can germinate and grow causing spoilage. Therefore, this microorganism was suggested as the target to be used in the design of adequate pasteurisation processes. The objectives of this work were to investigate the influence of temperature (T: 85-97 degrees C), total soluble solids (SS: 5-60 degrees Brix or % by weight) and pH (2.5-6.0) on D-values (decimal reduction time) of Alicyclobacillus acidoterrestris (type strain, NCIMB 13137) spores, and to fit a model using response surface methodology. A central composite face-centred experimental design was used, and the response, D-value determined in malt extract broth, ranged between 0.498+/-0.045 and 94.9+/-6.7 min. Within the factor ranges studied, temperature was the parameter that most affected the D-value. Following this was the SS and, lastly, the pH value. A linear decrease in D-value was observed with decreasing SS and pH, and a non-linear decrease in D-value was noticed with increasing temperature. A second order polynomial was successfully fitted to the data (R2 = 0.98). In general, D-values measured in real fruit systems, such as orange, apple and grape juices, blackcurrant concentrates, cupua?u (exotic fruit) extract and orange juice drink, were higher than those predicted by the malt extract broth model. This result emphasises the importance of experimental validation of any model-derived process.     

Fourier transform infrared spectroscopy, detection and identification of Escherichia coli O157:H7 and Alicyclobacillus strains in apple juice

Fourier Transform Infrared (FT-IR) spectroscopy (4000-400 cm(-1)) combined with multivariate statistical methods were used to identify and detect Escherichia coli O157:H7 from Alicyclobacillus spp. recovered from apple juice. Four treatments and a control in triplicate experiments (N=3) were studied; the first three treatments of pasteurized apple juice were inoculated with E. coli O157:H7 ATCC 35150, Alicyclobacillus acidoterrestris 1016 and Alicyclobacillus spp. C-Fugi-6 respectively. The fourth treatment was a 1:1 (v:v) mixed culture of both A. acidoterrestris 1016 and Alicyclobacillus spp. C-Fugi-6. The control was uninoculated pasteurized apple juice. The second derivative transformation and loadings plot over the range of 1800-900 cm(-1) highlighted the most distinctive variations among bacterial spectra. Loadings 1 and 2 were distinctively representative of the bacterial spectral data and accounted for 73% of the total variability. Treatments were noticeably segregated with distinct clustering by principal component analysis (PCA). Using soft independent modeling of class analogy (SIMCA) analysis, 88.3% of (E. coli O157:H7 ATCC 35150) spectra, 75.0% of (A. acidoterrestris 1016) spectra, 88.3% of (Alicyclobacillus spp. C-Fuji-6) spectra, and 80.0% of the mixed culture of both Alicyclobacillus strains spectra were correctly classified. Using the spectral features of bacterial cellular constituents such as nucleic acids, proteins, phospholipids, peptidoglycan, and lipopolysaccharides from examined bacterial cells, pure and mixed cultures of Alicyclobacillus spp. cells, and the pathogenic E. coli cells could be detected in apple juice.     

EFFECT OF ULTRASONIC TREATMENTS ON THERMOACIDOPHILIC ALICYCLOBACILLUS ACIDOTERRESTRIS IN APPLE JUICE

The effect of ultrasonic treatments on Alicyclobacillus acidoterrestris in apple juice was investigated. In general, inactivation of the cells was more pronounced at an elevated power level and as the processing time increased. Approximately 60% of the cells were inactivated after treating the apple juice with 300-W ultrasound for 30 min. The reduction reached more than 80% when the juice was processed for 60 min. The linear inactivation rates ( D values) of ultrasound on A. acidoterrestris were both process- and strain-dependent. The lowest D value at 36.18 min was found when using 600-W ultrasound to treat the A30 strain isolated from the air of a commercial apple juice processing plant, whereas the strain isolated from apple juice concentrate was found most resistant against ultrasound. Changes of sugar content, acidity, haze and juice browning were noted after ultrasonic treatments but did not adversely alter the juice quality.

PRACTICAL APPLICATIONS

Characterization of the effects of ultrasonic treatments on thermoacidophilic Alicyclobacillus acidoterrestris provides better understanding of how ultrasound technology could assist in alleviating the hard-to-detect spoilage caused by this spore former. Although ultrasonic technology might not be suitable as a standalone unit operation to inactivate high cell loads, the process could be integrated with other thermal or nonthermal technologies to enhance processing efficiencies, consequently preventing microbial spoilage caused by A. acidoterrestris .     

微波提取苹果汁中脂环酸芽孢杆菌DNA的PCR检测方法研究

根据Genbank公布的酸土脂环酸芽孢杆菌(Alicyclobacillus acidoterrestris DSM 3922T)鲨烯环化酶序列自行设计一对引物,利用微波技术直接从果汁样品中提取目标菌DNA,对引物特异性、微波法提取DNA的扩增效果及检测灵敏度进行探讨。结果表明：微波功率1000W、处理时间30s、经5000r/min离心2min即可获得酸土脂环酸芽孢杆菌基因组DNA,所得模板质量符合聚合酶链式反应检测要求,目的条带清晰,检测时间仅为2h,检测限为200CFU/mL,有望真正应用于苹果汁生产的在线检测。