青花椒精油抑制蜡样芽孢杆菌的活性与机理

青花椒精油是一种兼具药用和调味功能的天然精油,已报道具有良好的抑菌活性,应用前景广泛。然而,其抑菌机制却鲜有报道,本研究以蜡样芽孢杆菌（Bacillus cereus）为研究对象,从理化、形态学和代谢水平解析青花椒精油的抑菌机制。结果表明,青花椒精油中包含多种已经报道的抑菌活性成分,其对蜡样芽孢杆菌的最小抑菌质量浓度（minimum inhibitory concentration,MIC）和最小杀菌质量浓度（minimum bactericidal concentration,MBC）分别为2.0、4.0 mg/mL。青花椒精油能破坏蜡样芽孢杆菌的细胞壁和细胞膜,导致细胞质渗漏。在亚抑菌质量浓度（0.5 MIC）下的青花椒精油能够诱导蜡样芽孢杆菌在代谢水平上的显著变化。生物信息学分析表明,青花椒精油对蜡样芽孢杆菌造成了细胞膜损伤、能量代谢障碍、氨基酸代谢紊乱等多方面的影响。本实验对青花椒精油抑制蜡样芽孢杆菌的机制进行探讨,有助于青花椒精油在食品安全领域的开发和应用。     

紫苏黄酮抗菌活性表征

该试验利用超声波辅助乙醇提取法对紫苏黄酮进行提取。分别以大肠杆菌、金黄色葡萄球菌、霉菌及酵母菌为供试菌,采用滤纸片法等研究方法,通过抑菌圈、最小抑菌浓度（minimum inhibitory concentration,MIC）、最低杀菌浓度（minimum bactericidal concentration,MBC）等指标测定,探究紫苏黄酮抗菌活性。在此基础上,进一步研究紫苏黄酮抗菌活性的pH值稳定性和热稳定性。结果表明,紫苏黄酮对这4种供试菌均有一定的抗菌效果,其对大肠杆菌的抗菌活性最强（MIC和MBC分别为0.625、1.3 mg/mL）,其次为金黄色葡萄球菌（MIC和MBC分别为1.25、2.5 mg/mL）,再次为酵母菌（MIC和MBC分别为2.5、4.8 mg/mL）和霉菌（MIC和MBC分别为2.5、5 mg/mL）。在pH值为3～7的范围内,提取液浓度在各菌的MIC浓度下,均能完全有效抑制4种微生物的生长。温度条件在100℃以下时,紫苏黄酮的抗菌能力不受影响,当温度上升至121℃及以上时,会对抗菌活性产生负面影响。     

黔产青花椒精油成分分析及抑菌、抗氧化活性研究

以黔产青花椒为原料，采用水蒸气蒸馏法提取精油，运用气相色谱-质谱法（GC-MS）分析化学成分，并以大肠杆菌、耻垢分岐杆菌、铜绿假单胞菌、金黄色葡萄球菌为试验菌种评价抑菌活性和以DPPH自由基清除能力为指标评价精油的抗氧化活性。结果表明，从黔产青花椒精油中共鉴定出10个成分，以烯烃类、醇类、酮类为主，占精油总量的99.51%，其中相对含量高于3%的分别为β-芳樟醇（91.10%）、D-柠檬烯（3.12%）；精油对耻垢分岐杆菌的最小抑菌浓度（MIC）为4%，对大肠杆菌、铜绿假单胞菌、金黄色葡萄球菌的最小抑菌浓度（MIC）为8%。精油的抗氧化活性随着青花椒精油浓度的增加而逐渐升高，当浓度在52.1～884.2 mg/mL范围内时，最高清除率为92.747%。水蒸气蒸馏提取的青花椒精油中β-芳樟醇含量最高，具有较好的抑菌作用和抗氧化活性。     

桉叶精油包埋前后抑菌性能及成分比较研究

以包埋前后的桉叶精油为研究对象,采用平板打孔法和气相扩散法分别测定其对供试菌的抑菌活性和气相最低抑菌浓度(minimum inhibitory concentration,MIC)、气相最低杀菌浓度(minimum bactericidal concentration,MBC),探讨桉叶精油包埋前后抑菌性能差异。并用气相色谱-质谱联用技术对比分析桉叶精油包埋前后的化学成分。结果表明:桉叶精油包埋前后对供试细菌、酵母菌和霉菌均有一定的抑制作用。对酵母菌的抑制作用最强,其次为细菌,对霉菌的抑制效果稍差。桉叶精油包埋前后对供试菌的气相MIC和MBC未有太大差异。桉叶精油包埋前后主要成分大体相同,均以萜烯类和醇类化合物为主。包埋后桉叶精油中部分萜烯类化合物的消失或相对含量的减少导致抑菌性能稍有下降,但气相MIC不变,仅对酿酒酵母和桔青霉的气相MBC稍有上升。     

草果提取物对大肠杆菌和沙门氏菌抑菌机理研究

文章研究了草果中所含抑菌物质对大肠杆菌和沙门氏菌的抑菌机理。通过最小抑菌浓度(minimum inhibitory concentration,MIC)、最小杀菌浓度(minimum bactericidal concentration,MBC)和生长曲线的测定研究草果抑菌提取物对大肠杆菌和沙门氏菌细胞生长的影响;通过扫描电子显微镜(scanning electron microscopy,SEM)观察细菌细胞微观形态结构的变化;通过测定胞外碱性磷酸酶(alkaline phosphatase,AKP)研究草果抑菌提取物对大肠杆菌和沙门氏菌细胞壁通透性的影响。结果表明,草果提取物对供试菌的生长具有明显抑制作用,大肠杆菌的MIC和MBC值均为0.625mg/mL,沙门氏菌的MIC和MBC值约为1.25mg/mL;草果提取物在作用供试菌8h后,对其细胞结构造成了破坏,使得细胞壁的通透性增加。     

混料设计优化孜然、花椒、芥子混合精油对4 种果蔬采后真菌的抑菌效果

本研究用水蒸馏法提取孜然、花椒、芥子精油,并对其成分作气相色谱-质谱联用分析;采用微量稀释法,琼脂纸片扩散法测定3?种精油对4?种果蔬真菌粉红单端孢（Trichothecium roseum）、扩展青霉（Penicillium expansum）、互隔交链孢（Alternaria alternata）、硫色镰刀菌（Fusarium sulphureum）的最小抑菌浓度（minimum inhibitory concentration,MIC）、最小杀菌浓度（minimum bactericidal concentration,MBC）和菌落直径,利用单纯型增强质心混料设计数学模型拟合最佳精油混料比例,并利用响应面和方差分析混和精油抑菌效果。结果表明提取得到的孜然精油、花椒精油、芥子精油含量分别为2.29、2.52、1.12 mL/g。孜然精油主要化学成分为枯茗醛（21.89%）、β-蒎烯（20.49%）、γ-萜品烯（18.37%）、4-丙-2-环己基-1,4-二烯-1-甲醛（10.98%）、p-伞花烃（8.89%）、4-丙-2-环己基-3-烯-1-甲醛（7.95%）;花椒精油主要化学成分为芳樟醇（15.05%）、双戊烯（11.79%）、乙酸芳樟酯（11.17%）、β-蒎烯（9.33%）、（－）-4-萜品醇（6.04%）、桉叶油醇（4.62%）;芥子精油的主要化学成分为烯丙基异硫氰酸酯（50.97%）、4-异硫氰基-1-丁烯（44.91%）。当孜然、花椒、芥子精油混料比例分别为36.11%、19.45%、44.44%时,对4 种真菌的抑菌效果最佳,此时在统计学上存在显著性交互作用（P＜0.05）,具有良好的协同性;通过菌落直径大小分析得出混合精油相比于孜然精油、花椒精油单独作用具有较强的抑菌效果。该研究表明混料设计法可有效优化不同精油联合使用时的组合比,以实现较低精油用量下较好的抑菌效果,为天然抑菌剂的减量使用提供方法指导。     

留兰香-有机盐复合纳米微乳体系构建及抑菌活性研究

采用气相色谱-质谱联用技术分析留兰香精油组成,发现其主要物质为氧化胡椒烯酮和柠檬烯,分别占精油含量的39.74%和32.56%。构建拟三元相图研究留兰香微乳体系,发现抗坏血酸钠和山梨酸钾对微乳相行为没有显著影响。通过最低抑菌浓度（minimum inhibitory concentration,MIC）、最低杀菌浓度（minimum bactericidal/fungicidal concentration,MBC/MFC）和杀菌动力学实验研究抑菌活性,最后通过扫描电子显微镜和细胞膜通透性实验探究体系抑菌机理。结果显示：留兰香微乳体系对大肠杆菌、金黄色葡萄球菌、酿酒酵母菌和拜耳酵母菌均有抑制作用,MIC介于8～32 μL/mL,MBC/MFC介于32～128 μL/mL;并且在MBC条件下,大肠杆菌和金黄色葡萄球菌分别在处理60、90 min后全部失活。在留兰香微乳体系作用下,大肠杆菌核酸迅速外泄,并在电子显微镜照片中发现菌体形态出现扭曲、褶皱,甚至完全破裂。说明体系通过破坏细胞膜,改变细胞膜的结构和功能,使细胞内溶物外泄,从而杀死菌体。     

荔枝木质精油体外抑菌活性及稳定性研究

采用牛津杯法测定荔枝木质精油抑菌活性,以气体扩散法测定荔枝木质精油对7种常见食源性致病菌（大肠杆菌、金黄色葡萄球菌、乙型副伤寒沙门氏菌、鼠伤寒沙门氏菌、肠炎沙门氏菌、单核细胞增生李斯特菌、枯草芽孢杆菌）、3种常见霉菌（黑曲霉、桔青霉、黄曲霉）的最小抑菌浓度（minimum inhibitory concentration,MIC）和最小杀菌浓度（minimum bactericidal concentration,MBC）。同时进行荔枝木质精油抑菌活性的酸碱稳定性、热稳定性、紫外稳定性研究。结果表明：荔枝木质精油对受试细菌和霉菌均有抑制效果,其中对受试的革兰氏阳性菌抑制效果优于革兰氏阴性菌;对大肠杆菌、鼠伤寒沙门氏菌、乙型副伤寒沙门氏菌、肠炎沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特菌、枯草芽孢杆菌、黑曲霉、黄曲霉和桔青霉的 MIC 分别为 20、5、10、40、5、5、2.5、40、80、40 μL/mL;荔枝木质精油具有良好的紫外稳定性;加热处理并未对其抑菌效果产生显著影响;酸性条件下精油抑菌效果优于中性条件。     

费菜黄酮对冷却猪肉腐败菌及品质的影响

通过二倍稀释法和牛津杯法确定了费菜黄酮对3种冷却猪肉腐败菌的最低抑菌浓度(minimum inhibitory concentration,MIC)、最低杀菌浓度(minimum bactericidal concentration,MBC)和抑菌活性,并研究了不同质量浓度费菜黄酮对猪肉的保鲜效果。结果表明,费菜黄酮对实验菌有不同程度的抑制作用,其中对于气单胞菌的抑制性最强,MIC和MBC分别为0.675、2.5 mg/mL,假单胞菌次之,MIC和MBC分别为1.25、5 mg/mL,乳酸菌抑制性最差,MIC和MBC分别为1.25、10 mg/mL。综合菌落总数和色差指标,选择2 mg/mL作为费菜黄酮对冷却猪肉保鲜的适宜质量浓度。在4℃贮藏条件下,黄酮处理组组织结构、pH值、持水力、挥发性盐基氮值和硫代巴比妥酸值各品质指标均优于对照组。     

驼乳源乳铁蛋白嵌合肽对口腔致龋菌抗菌作用的初步探究

该研究旨在评估驼乳源乳铁蛋白(lactoferrampin-lactoferricin, LFA-LFC)嵌合肽对变异链球菌、唾液链球菌和远缘链球菌的抗菌效果并初步探究其作用机制。首先预测并鉴定驼乳源LFA-LFC嵌合肽的分子特性和二级结构；通过测定嵌合肽最低抑菌(minimum inhibitory concentration, MIC)、杀菌浓度(minimum bactericidal concentration, MBC),绘制时间-致死曲线，评估嵌合肽的抗菌效果；分析嵌合肽对生物膜的预防、消除作用。采用电子显微镜观察嵌合肽对细菌的作用位点，测定菌体DNA迁移率，对嵌合肽的抗菌机制进行初步研究。结果表明，驼乳源LFA-LFC嵌合肽对变异链球菌、唾液链球菌和远缘链球菌的最低抑菌浓度分别为32、32、64μmol/L;最低杀菌浓度分别为128、128、256μmol/L;细菌在1×MIC嵌合肽浓度处理30 min后完全致死；当嵌合肽浓度为128μmol/L时预防生物膜生成约80%,512μmol/L时消除生物膜约50%;4×MIC嵌合肽浓度处理细菌后发现其细胞膜均出现裂解，DNA未...     

葡萄籽和苹果原花青素对变形链球菌和金黄色葡萄球菌的抑制作用

研究苹果原花青素和葡萄籽原花青素对变形链球菌(Streptococcus mutans)和金黄色葡萄球菌(Staphylococcus aureus)的抑制作用,采用试管液体稀释法和琼脂稀释法抑菌实验测定其最低抑菌质量浓度(MIC)和质量最低杀菌质量浓度(MBC),并通过测定培养基pH值的变化,研究葡萄籽原花青素对变形链球菌产酸能力的影响。结果表明：苹果原花青素、葡萄籽原花青素对变形链球菌和金黄色葡萄球菌生长都有抑制作用;苹果原花青素对变形链球菌的MIC和MBC分别为1.25、4.00mg/mL,葡萄籽原花青素对变形链球菌的MIC和MBC分别为1.75、2.00mg/mL;苹果原花青素对金黄色葡萄球菌的MIC和MBC分别为4.00、9.00mg/mL,葡萄籽原花青素对金黄色葡萄球菌的MIC和MBC分别为2.00、6.50mg/mL。葡萄籽原花青素质量浓度大于1.00mg/mL对变形链球菌的产酸能力有显著抑制效果。     

抑制口腔变异链球菌的乳酸菌筛选及其抑菌机理

利用乳酸菌能抑制有害菌的特性,从实验室保藏的乳酸菌中筛选能抑制口腔变异链球菌的菌株,并测定了筛选菌株的最小抑菌浓度（minimal inhibitory concentration,MIC）,以及其对变异链球菌菌膜形成和黏附性的影响,初步研究其抑菌机理。结果表明,有5 株植物乳杆菌（Lactobacillus plantarum） K25、SKT109、YW3-2、YW5-1、YW1-1对变异链球菌具有明显的抑制作用。4 株变异链球菌指示菌中,变异链球菌（Streptococcus mutans）1.2499、1.2500对植物乳杆菌比较敏感。植物乳杆菌K25对变异链球菌的抑制作用比其他4株植物乳杆菌的作用更明显,其MIC最低（1.25×107～2.50×107 CFU/mL）。同时,菌株K25对变异链球菌菌膜形成的抑制率较高,还显著地降低变异链球菌的黏附率。对菌株K25抑菌机理的初步研究表明,其代谢过程中产生的乳酸和过氧化氢起到了主要的抑菌作用。     

不同低聚糖对变异链球菌、牙龈卟啉单胞菌和具核梭杆菌作用的体外研究

为更好地了解功能性低聚糖对口腔微生物的影响,本文采用体外实验方法,研究不同低聚糖对变异链球菌、牙龈卟啉单胞菌和具核梭杆菌生长、产酸、粘附性和生物被膜形成量等的影响。胰蛋白胨大豆肉汤（Trypticase Soy Broth,TSB）培养基中加入不同浓度的低聚半乳糖、低聚木糖、低聚果糖和低聚异麦芽糖制备培养基培养变异链球菌、牙龈卟啉单胞菌和具核梭杆菌,测定细菌在培养基中48 h生长过程中的OD值、pH、黏附能力及生物被膜形成量等指标。比较分析不同低聚糖对细菌发酵前后生长、产酸及黏附能力等特性的影响。结果表明:四种低聚糖对三种病原菌均有不同程度的抑制作用。其中,低聚木糖对三种病原菌的生长抑制效果最强;浓度为0.5%的低聚木糖和低聚半乳糖即能明显抑制三种菌液的产酸能力;相比糖醇和其它低聚糖,低聚木糖浓度≥5%时,对三种病原菌的抑制率均大于60%;7%浓度的四种低聚糖对三种病原菌的黏附率均<50%;低聚半乳糖和低聚木糖浓度>5%时,对三种病原菌生物被膜的形成具有较好的抑制作用。与山梨糖醇相比,复配糖（低聚木糖、木糖醇、低聚半乳糖）对三种病原菌的生长抑制作用更为明显。     

An in vitro study on the dental caries preventing effect of oligomeric procyanidins in sorghum episperm

Procyanidins are abundant in sorghum episperm. In this study, the effects of procyanidins from Sorghum bicolor (L.) moench episperm (sorghum procyanidins, SPC) in preventing dental caries were investigated through analysis on SPCs’ effects on cariogenic bacteria and ability in scavenging superoxide anion free radicals. In addition, the SPC with activity in preventing dental caries were identified. The ESI-MS/MS analysis results revealed that the compositions of SPC purified by macroporous resin were dimmers, trimers, tetramers, pentamers and hexamers of procyanidins and catechin/epicatechin monomers. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of SPC on Streptococcus mutans (Ingbritt C) and Streptococcus sobrinus 6715 were 8 and 16 mg/ml, respectively. The SPC had a remarkable inhibition effect (P < 0.05) on acid production of S. sobrinus (Ingbritt C) and S. mutans 6715 compared with the control group, and a dose-dependent relationship was observed. By increasing the concentration of SPC in the experimental condition (200–600 mg/L), the scavenging ratio of superoxide anion radicals increased significantly, indicating that SPC could be used as a potential dental-caries-preventative medicine with its antibacterial and antioxidant ability.     

A Lipoprotein-derived Antimicrobial Factor from Hen-egg Yolk is Active Against Streptococcus Species

ABSTRACT: Oral administration of hen-egg yolk provides protection against specific pathogens. We examined the antibacterial activity of fractionated egg yolk against 2 pathogenic Streptococcus strains, using an in vitro assay. A water-soluble protein fraction (WSPF) of egg yolk consistently inhibited the growth of S. mutans by 25%. The WSPF treated with pancreatin demonstrated > 80% inhibition of bacterial growth. Growth of S. sanguis was completely inhibited. Gel filtration and ion exchange chromatography established that anti-Streptococcal activity resided with lipoproteins. Antibacterial activity was released by crude lipase or a combination of lipase and protease treatment of egg lipoproteins. Thus, hen-egg yolk lipoproteins are important molecules for lipid-mediated antimicrobial activity.     

Antimicrobial effect of rosemary extracts

A rosemary extract commercially exploited (Oxy'less) as an antioxidant of lipids in foods was dissolved in ethanol (100 mg/ml), and the solution was tested against foodborne microorganisms. For gram-positive bacteria, the MIC of the ethanolic solution was 1% for Leuconostoc mesenteroides, 0.5% for Listeria monocytogenes, 0.5% for Staphylococcus aureus, 0.13% for Streptococcus mutans, and 0.06% for Bacillus cereus. It slowed the growth of Penicillium roquefortii and Botrytis cinerea. Up to 1% of the ethanolic solution had no activity on the gram-negative bacteria Escherichia coli, Salmonella Enteritidis, and Erwinia carotovora and on the yeasts Rhodotorula glutinis and Cryptococcus laurentii. Antibacterial activity of the rosemary extract was strongly influenced by the composition of the media. The MIC was reduced by low pH, high NaCl contents, and low temperatures. Low pH and high NaCl concentration had a synergistic effect on the MIC of the rosemary extract for S. aureus. Lipids, surface-active agents, and some proteins decreased its antibacterial activity, whereas pectin had no effect. The inhibitory effect was little modified by heat treatment (100 degrees C). The natural microflora of pasteurized zucchini broth was inhibited by 0.5% of the rosemary extract. The antibacterial activity was linked to the compounds extracted with hexane, which are presumably phenolic diterpenoids.     

罗伊氏菌素与儿茶素协同对变异链球菌的抑制作用

为探讨儿茶素与罗伊氏菌素（主要抗菌成分为3-羟基丙醛（3-hydroxypropionaldehyde，3-HPA））对诱发龋齿的变异链球菌的抑制作用及其机制，本研究测定了罗伊氏菌素与儿茶素单独使用和联合作用时对变异链球菌的最小抑菌浓度（minimum inhibitory concentration，MIC），经计算分级抑制浓度指数得知，当儿茶素（1/8 MIC）和3-HPA（1/4 MIC）联合使用时具有协同作用。测定生物膜含量、胞外多糖（extracellular polysaccharide，EPS）含量和有机酸含量3 个指标时发现，协同组使各指标含量显著减小（P＜0.05），且效果优于单一组。扫描电子显微镜结果显示，协同组生物膜结构改变明显、膜厚度降低、菌体数量减少。综上可看出，儿茶素和3-HPA协同使用能够抑制菌体的聚集与黏附、参与底物代谢，提高酸度、延缓生物膜生长成熟，可作为新型抑菌剂应用于防龋产品，为今后开发天然、安全的防龋产品提供理论依据。     

Essential oil of Curcuma longa inhibits Streptococcus mutans biofilm formation

Curcuma longa (C. longa) has been used as a spice in foods and as an antimicrobial in Oriental medicine. In this study, we evaluated the inhibitory effects of an essential oil isolated from C. longa on the cariogenic properties of Streptococcus mutans (S. mutans), which is an important bacterium in dental plaque and dental caries formation. First, the inhibitory effects of C. longa essential oil on the growth and acid production of S. mutans were tested. Next, the effect of C. longa essential oil on adhesion to saliva-coated hydroxyapatite beads (S-HAs) was investigated. C. longa essential oil inhibited the growth and acid production of S. mutans at concentrations from 0.5 to 4 mg/mL. The essential oil also exhibited significant inhibition of S. mutans adherence to S-HAs at concentrations higher than 0.5 mg/mL. S. mutans biofilm formation was determined by scanning electron microscopy (SEM) and safranin staining. The essential oil of C. longa inhibited the formation of S. mutans biofilms at concentrations higher than 0.5 mg/mL. The components of C. longa essential oil were then analyzed by GC and GC-MS, and the major components were α-turmerone (35.59%), germacrone (19.02%), α-zingiberene (8.74%), αr-turmerone (6.31%), trans-β-elemenone (5.65%), curlone (5.45%), and β-sesquiphellandrene (4.73%). These results suggest that C. longa may inhibit the cariogenic properties of S. mutans.     

茶黄素抑菌作用的研究

通过体外试验研究茶黄素(纯度40％)对大肠杆菌、金黄色葡萄球菌、黑曲霉、变形链球菌和远缘链球菌的抑菌作用.采用牛津杯测定法及最低抑菌浓度(MIC)测定法研究茶黄素的抑菌作用及对变链菌产酸能力的影响.结果表明,茶黄素对大肠杆菌、金黄色葡萄球菌有较强的抑菌作用,并与其浓度呈正相关,而对黑曲霉无抑菌作用;荼黄素能明显抑制口腔中主要的致龋菌变形链球菌和远缘链球菌的生长及产酸(P＜0.05),提高菌液pH值,具有一定的防龋效果.荼黄素是一种天然抗菌剂,作为食品添加剂可以提高食品的安全、卫生.     

In vitro Antilisterial Effects of Citrus Oil Fractions in Combination with Organic Acids

ABSTRACT:  The objectives of this study were to screen activity of citrus essential oil fractions (EOs) alone and in combination with organic acids against 2 species of Listeria . Five citrus EOs were initially screened by disc diffusion assay for antibacterial activity. Cold pressed terpeneless Valencia orange oil (CP terpeneless oil) had the strongest bacteriostatic (MIC) and bactericidal (MBC) properties at 0.55% and 1.67%, respectively. Four organic acids were tested for effectiveness against Listeria . Citric and malic acids proved to be the most effective with MBC of 1.1% alone. Assays were conducted to determine synergistic effects of EOs and citric or malic acids. There was a significant decrease in MIC and MBC to 0.04% EO plus 0.12% malic or citric acid. EOs from citrus paired with organic acids offer the potential as an all-natural antimicrobial for improving the safety of all-natural foods.