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1.
目的分离筛选出一株对甲胺磷敏感的双歧杆菌菌株。方法双歧杆菌菌株传代筛选,取一系列浓度(4.0、2.0、1.0、0.5和0.25μg/m L)的甲胺磷农药标准溶液30μL加入含TPY液体培养基的检测管溶液内,双歧杆菌菌株接种到检测管内,37℃厌氧培养12 h,测定检测管液体A值。结果筛选一株甲胺磷敏感的短双歧杆菌菌株LJM-006,微生物抑制法的最佳接种浓度107CFU/m L,最低检测限0.5 mg/L。结论筛选一株甲胺磷敏感的双歧杆菌LJM-006菌株,并可作为微生物法检测甲胺磷残留的菌株。  相似文献   

2.
目的分离筛选出一株对甲胺磷敏感的双歧杆菌菌株。方法双歧杆菌菌株传代筛选,取一系列浓度(4.0、2.0、1.0、0.5和0.25μg/mL)的甲胺磷农药标准溶液30μL加入含TPY液体培养基的检测管溶液内,双歧杆菌菌株接种到检测管内,37℃厌氧培养12h,测定检测管液体吸光度(A)值。结果筛选一株甲胺磷敏感的短双歧杆菌菌株LJM-006,微生物抑制法的最佳接种浓度为107 CFU/mL,最低检测限为0.5mg/L。结论筛选出一株甲胺磷敏感的双歧杆菌LJM-006菌株,并可作为微生物法检测甲胺磷残留的菌株。  相似文献   

3.
双歧杆菌原生质体的制备与回复研究   总被引:6,自引:0,他引:6  
进行了双歧杆菌原生质体的制备与回复相关技术研究 ,为其基因操作及相关研究提供技术基础。采用浓度分别为 1 ,5 ,1 0mg/LMutanolysin(变溶菌素 )对长双歧杆菌进行脱壁处理 ,以探讨其原生质体形成与时间和酶浓度的关系 ,然后选用较适宜的酶浓度 ( 5mg/LMutanolysin)制备其原生质体 ,并将其倾入自制的双层再生培养基上 ,观察其在不同环境条件下培养时的回复生长情况。结果表明 ,长双歧杆菌的细胞壁对Mu tanolysin较为敏感 ,用浓度为 5mg/L的Mutanolysin处理长双歧杆菌 40min ,在普通光学显微镜下即可见90 %的原生质体形成 ,当Mutanolysin浓度为 1 0mg/L时 ,只需 2 5min其原生质体形成率就达此值。制备的长双歧杆菌原生质体倾入自制的双层再生培养基中 ,在厌氧条件下能很好地回复生长。  相似文献   

4.
为研究荷叶黄酮对乳酸杆菌和双歧杆菌抗氧化能力的影响,0.25 mg/m L的荷叶黄酮添加至培养基培养保加利亚乳杆菌(Lactobacillus bulgaricus A105)、动物双歧杆菌(Bifidobacterium animalis B211),并提取细胞裂解液,依次从自由基清除能力、抗脂质过氧化能力及抗氧化物质活性三方面评估其抗氧化能力。结果显示,0.25mg/m L的荷叶黄酮提高了动物双歧杆菌(54.9%)及保加利亚乳杆菌(14.8%)对O-·2清除能力;其对保加利亚乳杆菌清除DPPH自由基能力稍有提升,对动物双歧杆菌清除DPPH自由基能力稍有抑制;其对保加利亚乳杆菌抗脂质过氧化能力提升率达658%,具有显著的协同增效作用。同时发现:经添加0.25 mg/m L的荷叶黄酮至培养基后,动物双歧杆菌和保加利亚乳杆菌胞内超氧化物歧化酶(SOD)酶活及谷胱甘肽(GSH)活力得到显著的提高。结果表明,荷叶黄酮的添加对保加利亚乳酸杆菌和动物双歧杆菌的抗氧化活性具有促进作用,且对保加利亚乳酸杆菌抗脂质过氧化能力具有明显的协同促进作用,可能与其胞内SOD酶活及GSH活力得到的显著的提升有关,具有潜在的应用价值。  相似文献   

5.
为研究荷叶黄酮对乳酸杆菌和双歧杆菌抗氧化能力的影响,0.25 mg/m L的荷叶黄酮添加至培养基培养保加利亚乳杆菌(Lactobacillus bulgaricus A105)、动物双歧杆菌(Bifidobacterium animalis B211),并提取细胞裂解液,依次从自由基清除能力、抗脂质过氧化能力及抗氧化物质活性三方面评估其抗氧化能力。结果显示,0.25mg/m L的荷叶黄酮提高了动物双歧杆菌(54.9%)及保加利亚乳杆菌(14.8%)对O-·2清除能力;其对保加利亚乳杆菌清除DPPH自由基能力稍有提升,对动物双歧杆菌清除DPPH自由基能力稍有抑制;其对保加利亚乳杆菌抗脂质过氧化能力提升率达658%,具有显著的协同增效作用。同时发现:经添加0.25 mg/m L的荷叶黄酮至培养基后,动物双歧杆菌和保加利亚乳杆菌胞内超氧化物歧化酶(SOD)酶活及谷胱甘肽(GSH)活力得到显著的提高。结果表明,荷叶黄酮的添加对保加利亚乳酸杆菌和动物双歧杆菌的抗氧化活性具有促进作用,且对保加利亚乳酸杆菌抗脂质过氧化能力具有明显的协同促进作用,可能与其胞内SOD酶活及GSH活力得到的显著的提升有关,具有潜在的应用价值。  相似文献   

6.
芽胞杆菌产生的环脂肽类生物表面活性素surfactin具有重要抗菌、促进生物膜形成等功能,其含量和同系物组分构成影响其功能。为了提高芽胞杆菌B006产生表面活性素的产量,通过单因素实验和正交实验,测定了碳源、氮源和无机盐等营养物质及接种量、培养时间、装液量和初始pH值对芽胞杆菌B006摇瓶发酵产生表面活性素的影响;采用排油圈法测定发酵液中表面活性素的产量,采用HPLC-MS方法比较培养基优化前后surfactin的产量和组分含量。结果表明,适合芽胞杆菌B006摇瓶发酵产生表面活性素的培养基组成为:牛肉膏10 g/L、玉米粉15 g/L、硝酸铵3 g/L和氯化钠3 g/L;适合的培养条件为:初始pH值7.0、接种量10%、装液量60 m L/500 m L,发酵周期64 h。优化后surfactin的产量约为314.73 mg/L,比优化前提高了74.88%;surfactin各组分比例发生改变,其中C16和C17组分的含量明显提高,为优化前相应组分含量的1.64和8.34倍。优化的培养基组分和培养条件可明显提高芽胞杆菌B006菌株产生surfactin的产量,改变surfactin同系物组分的构成,为利用芽胞杆菌B006进行高活性表面活性素的工业生产和代谢调控研究奠定了基础。  相似文献   

7.
替加环素与多粘菌素B对泛耐药鲍曼不动杆菌体外研究   总被引:2,自引:0,他引:2  
目的为治愈耐舒普深的泛耐药鲍曼不动杆菌(PDR-Ab)提供新药。方法替加环素采用二倍琼脂稀释法,多粘菌素B用E-test条法测分离目标菌株100株的最低抑菌浓度(M IC),并用WHONET 5.4软件分析数据。结果多粘菌素B对耐舒普深的泛耐药鲍曼不动杆菌株的M IC值分布情况为:1.5 mg/L为2株,1.0 mg/L为9株,0.75 mg/L为9株,0.5 mg/L为38株,0.38 mg/L为34株,0.25 mg/L为8株,均为敏感;替加环素对耐舒普深的泛耐药鲍曼不动杆菌株的M IC值分布情况为:≥32 mg/L为0株、16 mg/L为2株、8 mg/L为3株、4 mg/L为4株、2 mg/L为6株、1 mg/L为9株、0.5 mg/L为30株、0.25 mg/L为33株、0.125 mg/L为9株、0.06 mg/L为2株、0.03 mg/L为2株,敏感率为95.0%,中敏率为3.0%,耐药率为2.0%。结论替加环素或多粘菌素B是目前对耐舒普深的泛耐药鲍曼不动杆菌最有效药物之一。  相似文献   

8.
【目的】研究微生物的碳氮共脱除特性及其关键影响因素。【方法】以乙酸为唯一碳源分离获得的碳氮共脱除菌株Y5为模式菌株,分析菌株Y5的16S r RNA基因序列、碳源和氮源去除动力学,以及碳源种类、碳氮比(C/N)、溶解氧浓度(DO)、温度和p H等影响效果。【结果】菌株Y5归属于粪产碱杆菌。与葡萄糖及多种有机酸相比,菌株Y5在以乙酸为唯一碳源的条件下具有较高的TOC和NH4+-N去除速率。在好氧条件下,当起始TOC浓度为1 000 mg/L,氨氮浓度为110 mg/L时,菌株Y5的NH4+-N、TOC和总氮(TN)去除率分别达99.54%、92.95%和86.55%,最大NH4+-N、TOC和TN去除速率分别为903.58、505.81和406.03 mg/(L·d)。【结论】粪产碱杆菌Y5在以乙酸为唯一碳源的条件下具有较强的碳氮共脱除能力,其最佳反应条件为:C/N=10,p H 7.0-8.0,溶氧6.20 mg/L,反应温度为30°C。  相似文献   

9.
目的 对孟鲁司特钠联合双歧杆菌四联活菌片治疗小儿过敏性紫癜的临床疗效进行分析。方法 选取2015年3月1日至2018年3月31日我院收治的96例过敏性紫癜患儿为研究对象,依据随机数字表法分为A组、B组、C组、D组,每组24例。A组患儿施以常规临床治疗,B组在A组基础上给予孟鲁司特钠治疗、C组在A组基础上给予双歧杆菌四联活菌片治疗、D组在A组基础上施以孟鲁司特钠联合双歧杆菌四联活菌片治疗,4组患儿持续用药至出院后2个月。结果 A组患儿治疗有效率为66.67%,皮肤紫癜消失时间为(10.49±3.01)d,关节肿痛消失时间为(5.87±1.56)d,腹痛消失时间为(4.05±1.28)d,Alb水平为(27.11±7.80)mg/L,β2-MG水平为(0.31±0.15)mg/L,治疗后复发率为29.17%。D组患儿治疗有效率为91.67%、皮肤紫癜消失时间为(6.98±2.57)d,关节肿痛消失时间为(3.07±1.04)d,腹痛消失时间为(2.77±1.05)d,Alb水平为(22.00±8.47)mg/L,β2-MG水平为(0.19±0.06)mg/L,治疗后复发率为4.17%。D组患儿各指标与A组比较差异均有统计学意义(均P<0.05)。B、C组患儿治疗有效率高于A组,但低于D组。B、C组患儿皮肤紫癜消失时间、关节肿痛消失时间、腹痛消失时间与A组比较差异无统计学意义(P>0.05),与D组相比差异有统计学意义(P<0.05)。结论 孟鲁司特钠联合双歧杆菌四联活菌片对小儿过敏性紫癜的疗效显著,能减少患儿不良反应发生,促进患儿恢复健康,值得临床推广。  相似文献   

10.
以红鸥羽衣甘蓝下胚轴为试材,首先优化了不定芽分化体系,其次探讨了农杆菌介导遗传转化过程中农杆菌侵染浓度、侵染时间对不定芽分化率的影响及不定芽分化和生根过程中潮霉素B筛选压力,最后对抗性植株进行了潮霉素B筛选基因的PCR检测。结果表明,在MS+6-BA 5.0 mg/L+Ag NO3 9.0 mg/L培养基中不定芽分化率最高,为84.17%;农杆菌侵染浓度OD600值为0.5、侵染5 min时利于遗传转化,分化率为69.17%;不定芽分化和生根过程中潮霉素B筛选压力分别为4.0 mg/L和30.0 mg/L;潮霉素B筛选基因的PCR鉴定结果,在预期的602 bp处出现了条带,初步证明潮霉素B筛选基因已整合到羽衣甘蓝基因组中。  相似文献   

11.
应用3天种群增长实验方法,在(25±1)℃、无光照、以3.0×106 cells/mL的斜生栅藻为轮虫的食物等条件下,研究了亚致死浓度(0.01、0.1、1.0、10.0、100.0、1000.0和10000.0 µg/L)的甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态的影响。结果显示,甲胺磷和乙酰甲胺磷显著地影响萼花臂尾轮虫的种群增长率、混交雌体数与非混交雌体数的比值和混交率。甲胺磷显著地影响萼花臂尾轮虫种群中的带卵雌体数与不带卵雌体数的比值,但乙酰甲胺磷对其无显著的影响。和对照组相比,浓度为100.0 µg/L 的甲胺磷和浓度为1.0-10,000.0 µg/L 的乙酰甲胺磷均使轮虫种群增长率显著增大,而浓度为1000.0 µg/L和10000.0 µg/L的甲胺磷却使之显著减小;1000.0 µg/L 的甲胺磷使轮虫种群中的带卵雌体数与不带卵雌体数的比值显著上升,0.1 µg/L的甲胺磷和10.0-10000.0 µg/L的乙酰甲胺磷均使轮虫种群中的混交雌体数与非混交雌体数的比值显著上升,0.1µg/L 的甲胺磷和10 µg/L的乙酰甲胺磷均使轮虫的混交率显著增大,10.0-0000.0 µg/L 的乙酰甲胺磷使轮虫休眠卵产量显著提高。上述结果表明,亚致死浓度的甲胺磷和乙酰甲胺磷对萼花臂尾轮虫实验种群动态具有显著的影响。  相似文献   

12.
目的 探索肠道益生菌在抗生素残留监测中的应用并建立牛奶中多种抗生素残留的微生物学筛检方法。方法 厌氧培养分离健康人肠道益生菌并用纸片扩散法检测其对四环素、红霉素、青霉素、庆大霉素、氯霉素和万古霉素的敏感性。根据药敏结果及产酸能力选择工作菌株。建立基于厌氧培养的益生菌抗生素残留筛检方法并优化实验条件,测定其对不同抗生素的检测限,通过测定加标模拟阳性和阴性样本计算方法的准确性。结果 短双歧杆菌A2可用于牛奶抗生素残留筛检。应用短双歧杆菌筛检牛奶中四环素、红霉素和青霉素的检测限分别为80 μg/L、40 μg/L和3 μg/L,检测加标四环素、红霉素和青霉素的牛奶样品准确性为100.0%、92.5%和90.0%,无假阴性结果。结论 应用益生菌的筛检方法敏感、简便,对环境和操作者友好,适用于监测牛奶中四环素、红霉素和青霉素等多种抗生素。  相似文献   

13.
It is believed that an important criterion for a potential probiotic strain is that it is capable of adhering to mucosal surfaces in the human gastrointestinal tract. The purpose of this study was to investigate a possible relationship between exopolysaccharide production and adhesion to Caco-2 cells by Bifidobacterium breve A28 and Bifidobacterium bifidum A10. In a preselection process, the hemagglutination abilities of these bacteria were determined prior to undertaking adhesion studies. B. breve A28, which produces large amounts of EPS (97.00 ± 2.00 mg/l) and has good hemagglutination abilities (+3) was found to adhere strongly to Caco-2 cells. Under gastrointestinal conditions, the high EPS producing- B. breve A28 was found to have better viability and adhesion to Caco-2 cells than the low EPS producing- B. bifidum A10. Also, B. breve A28 was found to be more effective at inhibiting Escherichia coli ATCC 11229 than B. bifidum A10. This investigation showed that high EPS production and adhesion ability may be important in the selection of bifidobacteria as probiotic strains.  相似文献   

14.
Quantitative real-time PCR assays targeting the groEL gene for the specific enumeration of 12 human fecal Bifidobacterium species were developed. The housekeeping gene groEL (HSP60 in eukaryotes) was used as a discriminative marker for the differentiation of Bifidobacterium adolescentis, B. angulatum, B. animalis, B. bifidum, B. breve, B. catenulatum, B. dentium, B. gallicum, B. longum, B. pseudocatenulatum, B. pseudolongum, and B. thermophilum. The bifidobacterial chromosome contains a single copy of the groEL gene, allowing the determination of the cell number by quantification of the groEL copy number. Real-time PCR assays were validated by comparing fecal samples spiked with known numbers of a given Bifidobacterium species. Independent of the Bifidobacterium species tested, the proportion of groEL copies recovered from fecal samples spiked with 5 to 9 log(10) cells/g feces was approximately 50%. The quantification limit was 5 to 6 log(10) groEL copies/g feces. The interassay variability was less than 10%, and variability between different DNA extractions was less than 23%. The method developed was applied to fecal samples from healthy adults and full-term breast-fed infants. Bifidobacterial diversity in both adults and infants was low, with mostly ≤3 Bifidobacterium species and B. longum frequently detected. The predominant species in infant and adult fecal samples were B. breve and B. adolescentis, respectively. It was possible to distinguish B. catenulatum and B. pseudocatenulatum. We conclude that the groEL gene is a suitable molecular marker for the specific and accurate quantification of human fecal Bifidobacterium species by real-time PCR.  相似文献   

15.
The molecular interactions between the bifidobacterial cell and its natural environment, namely, the gastrointestinal tract of its host, are particularly important in understanding the presumed positive effects of Bifidobacterium on the health status of the host. In this study an export-specific reporter system, designed for use in gram-positive organisms and based on the use of the staphylococcal nuclease (Nuc) as a reporter, was employed to identify exported proteins in Bifidobacterium breve UCC2003. A B. breve genomic library of translational fusions to the Nuc-encoding gene devoid of its own export signal was established in the shuttle vector pFUN (I. Poquet, S. D. Ehrlich, and A. Gruss, J. Bacteriol. 180:1904-1912, 1998) and screened for bifidobacterial export signals. Sequence analysis of the fusion proteins obtained that displayed a nuclease-producing phenotype in both Lactococcus lactis and B. breve predicted the presence of a classical signal peptide and/or single or multiple transmembrane domains, thus indicating that some of the export signals in B. breve are comparable to those used in L. lactis. Cell fractionation studies, zymograms, nuclease assays, and Western blotting were employed to confirm the function of the predicted signals and to determine the location and activity of the exported fusion proteins in B. breve and/or L. lactis.  相似文献   

16.
To investigate the effects of administration of raffinose and encapsulated Bifidobacterium breve JCM 1192T cells on the rat cecal microbiota, in a preclinical synbiotic study groups of male WKAH/Hkm Slc rats were fed for 3 weeks with four different test diets: basal diet (group BD), basal diet supplemented with raffinose (group RAF), basal diet supplemented with encapsulated B. breve (group CB), and basal diet supplemented with both raffinose and encapsulated B. breve (group RCB). The bacterial populations in cecal samples were determined by fluorescence in situ hybridization (FISH) and terminal restriction fragment length polymorphism (T-RFLP). B. breve cells were detected only in the RCB group and accounted for about 6.3% of the total cells as determined by FISH analysis. B. breve was also detected only in the RCB group by T-RFLP analysis. This was in contrast to the CB group, in which no B. breve signals were detected by either FISH or T-RFLP. Increases in the sizes of the populations of Bifidobacterium animalis, a Bifidobacterium indigenous to the rat, were observed in the RAF and RCB groups. Principal-component analysis of T-RFLP results revealed significant alterations in the bacterial populations of rats in the RAF and RCB groups; the population in the CB group was similar to that in the control group (group BD). To the best of our knowledge, these results provide the first clear picture of the changes in the rat cecal microbiota in response to synbiotic administration.  相似文献   

17.
Fecal microbiota of 31 breast-fed, 26 mix-fed, and 11 bottle-fed infants were analyzed by using terminal restriction fragment length polymorphism (T-RFLP), and culture method. We first determined the total and cultivated bacterial counts in infant fecal microbiota. Only approximately 30% of bacteria present in fecal microbiota were cultivable while the remainder was yet-to-be cultured bacteria. Sixty-eight fecal samples were divided into two clusters (I and II) by T-RFLP analysis, and then subdivided into five subclusters (Ia, Ib, IIa, IIb and IIc). There was no clear relationship between clusters and feeding method. A proportion of bifidobacteria was detected in the fecal material by PCR method using species-specific primers. The predominant Bifidobacterium spp. was Bifidobacterium longum longum type (43 samples (63.2%)), followed by B. longum infantis type (23 samples (33.8%)) and B. breve (16 samples (23.5%)). The distribution of Bifidobacterium spp. was similar in the three feeding groups. In contrast, the high incidence of B. breve in cluster I, especially subcluster Ia and B. longum longum type in cluster II, especially subcluster IIa and IIc were characterized by T-RFLP method. Our results showed that the colonization of Bifidobacterium spp. in infant feces correlated with the T-RFLP clusters.  相似文献   

18.
The production of an intracellular secondary metabolite rosmarinic acid (RA) by plant cell suspensions of Anchusa officinalis cultivated with intermittent medium exchange is investigated. Initially, a two-stage perfusion culture method was employed. After being cultured in the batch mode for ca. 6 days in B5 medium plus 3% sucrose, 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.1 mg/L kinetin (2,4-D B5 medium), Anchusa culture was cultivated to high cell density by perfusion during the growth stage using a hormone-free Gamborg B5 medium supplemented with 6% sucrose. This was followed by a production stage, in which a complete medium exchange into B5 medium plus 3% sucrose and 0.25 mg/L naphthleneacetic acid (NAA) was conducted. The two-stage perfusion culture had a higher maximum culture RA concentration but a lower RA content per cell than the batch stock culture maintained in the 2,4-D B5 medium. Higher culture RA concentration was due primarily to high cell density. The high packed cell volume, however, seemed to reduce the synergistic effect of NAA on RA synthesis. Subsequently, a single-stage perfusion culture method was investigated. The best result was obtained by growing the culture in the batch mode for ca. 10 days using B5 medium supplemented with 3% sucrose and 0.25 mg/L NAA, followed by perfusing the culture with B5 medium plus 6% sucrose and 0.25 mg/L NAA at a constant perfusion rate of 0.1/day. A maximum cell dry weight of 35 g/L and a RA concentration of almost 4 g/L were achieved. This is the highest RA concentration ever reported in the Anchusa culture. (c) 1993 John Wiley & Sons, Inc.  相似文献   

19.
Probiotics are used for the improvement of gut disorders. To explore the potential of probiotics, a gnotobiotic study using BALB/c mice to analyze epithelial gene expression was performed. Microarray analysis of probiotic strain-monoassociated mice showed that Lactobacillus casei Shirota and Bifidobacterium breve Yakult noticeably affected gene expression in the ileal and colonic epithelial cells, respectively, although to a smaller extent than segmented filamentous bacteria (SFB). Lactobacillus casei Shirota enhanced the gene expression involving defense/immune functions and lipid metabolism more strongly than B. breve Yakult. In the colon, expression of a chloride transporter was slightly enhanced, although downregulation of many genes, such as guanine nucleotide-binding protein, was evident in mice with B. breve Yakult compared with the ones with L. casei Shirota. SFB affected gene expression more strongly than the probiotic strains. In particular, alpha(1-2) fucosyltransferase and pancreatitis-associated protein were significantly enhanced only in SFB-monoassociated mice but not probiotic strain-monoassociated mice. Gene expression of SFB-monoassociated mice was either stimulated or repressed in a manner similar to or opposite that of conventional colonized mice. Taken together, probiotic strains of L. casei Shirota and B. breve Yakult differentially affect epithelial gene expression in the small intestine and colon, respectively.  相似文献   

20.
Bifidobacteria are nonpathogenic, anaerobic domestic bacteria with health-promoting properties for the host. In our previous study, Bifidobacterium longum (B. longum) were found to be localized selectively and to proliferate within solid tumors after systemic application. Additionally, B. longum transformed by shuttle-plasmid including the cytosine deaminase (CD) gene expressed active CD, converted the prodrug 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU). We also demonstrated antitumor efficacy with a transformant of B. longum in rats. In this study, we found that Bifidobacterium breve (B. breve), the smallest species of human-derived bifidobacterium, expressed the exogenous transgene (CD), that CD enzymatic activity in the transformant of B. breve was much higher, and that the segregational stability of the plasmid was greater than that of B. longum. Thus, numerous transformants of B. breve were detected solely in the tumors after systemic administration. We consider the transformant of B. breve to be more beneficial in our enzyme/prodrug therapy.  相似文献   

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