短双歧杆菌增殖培养基的优化研究

本研究针对短双歧杆菌的营养需要,采用正交实验设计优化增殖培养基,结果表明,酵母膏是短双歧杆菌的显著影响因素, 最佳的优化培养基配比是酵母膏0.9%,胰蛋白胨0.6%,大豆蛋白胨0.6%,葡萄糖0.6%,双歧因子0.2%,生长因子10%.用经优化的增殖培养基配方测定短双歧杆菌的生长曲线及其pH的变化,确定该菌增殖培养的最适终止时间约为18h,此时平板菌落计数的菌数可高达1.5×1010cfu/ml.     

响应面法优化青春双歧杆菌增殖培养基

为提高青春双歧杆菌在发酵液中的活菌数,以MRS为培养基,采用响应面法对培养基进行优化,同时比较了优化前后的生长曲线与pH的变化。通过响应面分析结果得到青春双歧杆菌的增殖培养基配方：葡萄糖15.85g/L、低聚果糖15.85 g/L、胰蛋白胨12.04 g/L、牛肉膏7.23 g/L、酵母粉9.63 g/L、柠檬酸铵2.75 g/L、K2HPO4·3H2O 2.75 g/L、乙酸钠6.875 g/L、吐温-80 1.0 mL、MgSO4·7H2O 0.2 g/L、MnSO4·H2O 0.05 g/L、L-半胱氨酸盐酸盐0.5 g/L。该菌在增殖培养基中28 h达到稳定期,比优化前缩短12 h;活菌数达到8.9×109 CFU/mL,是优化前的1.73倍。     

纳他霉素融合子Sr-1发酵培养基成分优化

本实验根据对融合子Sr-1菌株摇瓶培养基组成及发酵条件单因素优化基础上,利用响应面法获得最佳的培养基配方为葡萄糖33g/L、糊精7.7g/L、胰蛋白胨26g/L、酵母浸粉8.2g/L、初始pH7.0、最佳接种龄28h,发酵周期72h。     

优化两歧双歧杆菌培养基的增菌研究

针对两歧双歧杆菌的营养需要,采用正交实验设计优化增殖培养基。结果表明,酵母膏是短双歧杆菌的显著影响因素。最佳的优化培养基配比是:酵母膏0.9%,胰蛋白胨0.3%,大豆蛋白胨0.3%,葡萄糖2.0%,双歧因子0.8%,生长因子10%(均为质量分数)。用经优化的增殖培养基配方测定两歧双歧杆菌的生长曲线及其pH值的变化,确定该菌增殖培养的最适终止时间约为18h,此时平板菌落计数的菌数可高达2.1×1010mL-1。     

罗伊氏乳杆菌LT018高密度培养生长因素的研究

罗伊氏乳杆菌LT018是经过筛选的源自巴马百岁老人粪便中的优良益生菌株。为优化出适合其高密度生长的培养基,本研究以TPY为基础培养基,采用单因素方法确定增值培养基的成分为胰蛋白胨、酵母粉、麦芽糖、柠檬酸、柠檬酸钠、西红柿汁和L-半胱氨酸。采用Plackett-Burman实验设计方法得出胰蛋白胨、柠檬酸和L-半胱氨酸对菌株LT018的生长影响显著,继而进行最陡爬坡实验,通过RSM确定显著因子的最优组合为:胰蛋白胨含量12.13 g/L,柠檬酸0.4 g/L,L-半胱氨酸0.6 g/L。在上述培养基的基础上,利用单因素方法确定培养最佳条件为:接种量为4%,温度为37℃,初始p H为7.0,静置培养。此时该菌株的活菌数可达到7.13×1010cfu/m L,是未优化前的10.7倍。     

薛氏丙酸杆菌产抑菌性代谢物发酵培养基优化

本文利用单因子实验和响应面实验设计对薛氏丙酸杆菌产抑菌性代谢物的发酵培养基组成进行优化,以降低培养基成本和进一步提高代谢物抑菌活性。实验结果表明,玉米浆和大豆蛋白胨可以代替原始培养基中的酵母提取物和胰蛋白胨,通过Box-Behnken实验设计和响应面分析法优化得到了薛氏丙酸杆菌产抑菌性代谢物的发酵培养基组成为葡萄糖8.2g/L,大豆蛋白胨5.1g/L,玉米浆12.7g/L,在此条件下,代谢物抑菌活性的理论值为29.5AU/m L,实际测定值为29.4AU/m L。优化之后,薛氏丙酸杆菌代谢物抑菌性提高了57.2%,培养基成本大大降低。     

响应面法优化豆乳链球菌增殖培养基

针对优良酸豆乳发酵菌株豆乳链球菌（Streptococcus sojalactis）SY1.1增殖培养基进行优化,研究碳氮源对豆乳链球菌生长的影响,并采用响应面法优化豆乳链球菌SY1.1的增殖培养基配方。结果表明：以AST为基础培养基,优选大豆蛋白胨和蔗糖为最适氮源和碳源,质量比为1∶2,采用Plackett-Burman设计对11 种促乳酸菌生长因子进行评价,利用中心旋转组合设计和响应面分析确定增殖培养基的配方为：蔗糖10 g/L、大豆蛋白胨20 g/L、磷酸氢二钾2 g/L、番茄汁85.8 mL/L、胡萝卜汁109 mL/L、玉米浆6 g/L,pH 6.8。SY1.1以接种量2%,在37 ℃培养12 h时,活菌数可达（8.9±0.2）×108 CFU/mL,较初始豆浆培养基的活菌数（1.03×108 CFU/mL）提高了7.64 倍。     

响应曲面法优化短双歧杆菌Bifidobacterium breve A04培养基

本文通过响应曲面法(RSM)达到优化双歧杆菌培养基组分的目的。以TPYG培养基为基础培养基,采用FFD(factionalfactorialdesign)实验设计法,筛选出三种最佳生长强化因子葡萄糖、酵母粉和初始pH值;通过快速登高法摸索出生长强化因子的最佳浓度范围为:葡萄糖5.0g/L,酵母膏3.01g/L以及初始pH6.0;利用旋转中心组分设计法(RCD)和响应曲面法(RSM)确定关键组分的最佳浓度并进行了验证。最终优化培养基组分为葡萄糖5.194g/L,酵母膏2.829g/L,低聚果糖2g/L,西红柿汁50ml,肝浸液5g/L,Tween801ml,半胱氨酸盐酸盐0.3g/L,pH6.463。最终优化出的培养基获得的菌体干重达到4.462g/L,比优化前3.16g/L提高了1.43倍。     

牛胎盘废弃物制备食品微生物培养基

采用废弃牛胎盘下脚料为氮源制备新型培养基。并采用Plackett-Burman法、最陡爬坡试验和响应面试验(Box-Behnken设计法)相结合的方法对大肠杆菌培养基进行优化。结果表明:牛胎盘下脚料经水解后,替代LB培养基中的酵母浸粉,显示出较高的生长水平。优化后的新型培养基(NTP)组成为:葡萄糖2.8 g/L,NaCl 11 g/L,胰蛋白胨10.3 g/L,牛胎盘水解物16.7 g/L,pH 7.85,最终得到活菌数量较高的培养基配方。     

植物乳杆菌LB-B1产细菌素发酵条件的优化

为提高分离自新疆传统发酵乳制品酸马奶中植物乳杆菌LB-B1产细菌素的能力,对其产细菌素的发酵条件进行了优化,分别研究了培养条件(时间、温度、培养基初始pH值)和培养基成分对细菌素产量的影响。通过单因素试验和正交试验,确定产植物乳杆菌LB-B1的最佳培养条件为37℃静置培养20h,培养基初始pH值为6~7;最佳培养基成分组合为葡萄糖30g/L、胰蛋白胨10g/L、牛肉膏10g/L、酵母粉5g/L、无水乙酸钠5g/L、柠檬酸铵2g/L、磷酸氢二钾2g/L、硫酸镁0.28g/L、硫酸锰0.25g/L、吐温-80 4mL/L。在优化发酵条件下,植物乳杆菌LB-B1产细菌素高达10240AU/mL,提高了3倍。     

A medium for the detection of yeasts using a conductimetric method

A new medium is described for the detection of yeasts using a Malthus microbial growth analyser which measures conductivity. This medium is superior to previous media devised for use in the Malthus, and allows the detection of a wide range of yeasts including those important in food spoilage. The new medium was inferior to existing media used to detect yeasts in the Bactometer 32 instrument which measures impedance.     

不同增菌液对单增李斯特菌的增菌效果比较

目的比较不同选择性增菌液对单增李斯特菌增菌效果及对干扰菌的抗干扰性。方法参考国标GB/T 4789.30-2016和国际ISO 11290-1-2017,将2种来源的单增李斯特菌(标准菌株、分离菌株)和2种干扰菌(大肠埃希氏菌、金黄色葡萄球菌)分别接种到4种不同增菌液中,观察增菌后菌液的生长情况。结果研究发现低浓度菌液(102数量级), Fraser肉汤对单增李斯特菌的增菌效果及抗干扰性优于LB肉汤。选择性添加剂浓度过高会抑制李斯特菌的生长,通过选择合适的选择性添加剂及改变添加剂的浓度可以提高检出率。结论在实验室日常检验过程中,需考虑样品类别,选择合适的培养基。提高检测的准确率。     

产弹性蛋白酶EL314CF10菌株种子培养基优化的研究初报

采用两水平部分因子实验设计 (FFD)研究了产弹性蛋白酶菌株的种子培养基各组分对菌体生长的影响 ,并找出主要影响因子为牛肉膏、酵母膏 ,二者都在 99%的概率水平上对菌体生长有正效应。而蛋白胨对菌体生长影响是负效应 ,但不显著。NaCl对菌体生长影响也极不显著。由方差分析结果可知 ,选取的水平接近最大响应区域。因此不再进行最陡爬坡实验。最后用中心复合设计及响应面分析确定主要影响因子的最佳浓度。由实验结果可知 ,菌株在优化后的种子培养基中生物量比未优化培养基高。     

Development of a chemically defined medium for studying foodborne bacterial–fungal interactions

There is a growing interest for using natural preservatives in the food and dairy industries including the application of bacterial cultures to inhibit fungal spoilage. Several antifungal metabolites from bacteria have been identified, but their relative importance has been difficult to establish. In dynamic systems such as fermented milk products, the complexity of the food matrix affects detection, identification and quantification of antifungal metabolites, and thereby the understanding of the bacterial–fungal interactions. To ease the identification and quantification of bacterial metabolites (as judged by ultra-performance liquid chromatography/mass spectrometry) a chemically defined interaction medium (CDIM) was developed. The medium supported growth of antifungal cultures such as Lactobacillus paracasei and Propionibacterium freudenreichii, as well as spoilage moulds and yeasts isolated from fermented milk products. Both strong and weak antifungal interactions observed in milk could be reproduced in CDIM. The medium seems suitable for studying antifungal activity of bacterial cultures.     

Determinants of bacterial replication rates in mastitic whey

Bacterial growth was measured by a turbidimetric microtechnique in the whey of milk samples from quarters of cows with subclinical mastitis. Samples were grouped according to bacterial isolates recovered and the effects of bacterial species and whey on bacterial growth rates were analysed. Different strains of bacteria and different whey samples gave highly significant differences in bacterial replication rates. Except for penicillin-resistant Staphylococcus aureus, bacteria grew better in whey from mastitic milk where the inflammation was caused by the same bacterial species than in other mastitic milk samples. Inflammation caused by major pathogens generally enhanced the growth in whey of any type of major pathogen. Since mastitis pathogens showed enhanced growth in whey prepared from the same milk from which they were isolated, specific antibacterial factors in the whey did not appear to restrict bacterial growth in whey. The nutritional quality of the medium seems to be the important determinant of bacterial growth.     

顶空气相色谱法快速检测卫生纸中的细菌含量

本文基于细菌在培养过程中的生长繁殖数量与其代谢产物(CO2)释放量的关系,建立了一种利用顶空气相色谱技术快速测定卫生纸中细菌含量的新方法;同时,考察了不同培养基、纸样处理及接种方式、培养液浆浓对检测速度及可靠性的影响。结果表明:采用TGEA培养基比LB液体培养基能获得更快的检测速度;在纸样处理及接种方式中I,SO 8784-1:2005(E)标准的方法优于GB/T 20810-2006标准;为了提高检测的灵敏度和检测的准确性,培养液的纸浆浓度不应高于0.5%(摇床转速360r/min)。在上述条件下,该方法在培养时间8h时即可实现对卫生纸细菌含量的准确检测,与传统方法相比大大提高了检测的效率。     

发酵肉制品中乳酸菌计数培养基的优化

采用平板计数法比较了5种乳酸菌在4种培养基(MRS、SL、TJA与酪蛋白山梨酸培养基)上的生长情况,选择乳酸菌生长最好的培养基,对其pH值、添加山梨酸和纳他霉素进行改良,检测了5种乳酸菌及6种杂菌在此培养基上的生长情况,并用发酵肉制品样品进行验证.结果表明,采用改良TJA培养基对发酵肉制品中乳酸菌计数效果最好,在pH 5.5、添加山梨酸(1000mg/L)和纳他霉素(20mg/L)条件下能排除其他杂菌的干扰,提高乳酸菌计数的准确性.优化后的改良TJA培养基能较好地排除发酵肉制品中的杂菌对乳酸菌计数的影响,可应用于发酵肉制品中乳酸菌的计数.     

3种不同培养基和1种测试片检测水溶性化妆品中菌落总数的结果分析

目的使用t检验法分析3种不同培养基和1种测试片检测水溶性化妆品中菌落总数结果。方法参照《化妆品安全技术规范》(2015年版),使用卵磷脂吐温80营养琼脂、TTC营养琼脂、平板计数琼脂方法进行样品菌落总数检测,然后使用Petrifilm TM菌落测试片检测样品。运用配对t检验法评价4种不同方法对水溶性化妆品中菌落总数计数结果一致性的影响。结果在10~2、10~3 CFU/m L 2种不同浓度,平板计数琼脂法结果波动性最大,无法抑制菌落蔓延,而TTC营养琼脂和菌落测试片相对卵磷脂吐温80营养琼脂,具有抑制菌落蔓延生长、提高计数结果准确性的作用。结论在一定浓度下,可根据实际情况来选择相应培养基检测水溶性化妆品中菌落总数以提高检测效率和质量。     

Automated turbidometry for predicting colony forming units in raw milk

The Bioscreen turbidometer was used to study bacterial growth in a laboratory medium inoculated with raw milk. The results showed that the number of colony forming bacteria in milk determined by the plate count method can be estimated from turbidometric parameters of bacterial growth. A formula predicting the bacterial count in raw milk was developed.     

响应面法优化产蛋白酶菌株的发酵条件

从新疆吐鲁番地区土样中分离筛选获得1株产蛋白酶菌株Bacillus tequilensis C9,采用Plackett-Burman(PB)设计法筛选出影响其产酶条件的主效应因素为葡萄糖含量、装液量和接种量.应用中心组合设计以及响应面分析,建立了以蛋白酶酶活为响应值的二次回归方程模型,得到最优发酵产酶条件为:葡萄糖含量1.3％,装液量45mL,接种量5％.此条件下蛋白酶活为86.17 U/mL,与优化前相比酶活提高2.1倍.