Antagonistic Potential against Pathogenic Microorganisms and Hydrogen Peroxide Production of Indigenous Lactobacilli Isolated from Vagina of Chinese Pregnant Women

Objective To investigate the indigenous lactobacilli from the vagina of pregnant women and to screen the isolates with antagonistic potential against pathogenic microorganisms. Methods The strains were isolated from pregnant women＇s vagina and identified using the API50CH system. The ability of the isolates to produce hydrogen peroxide was analyzed semi-quantitatively using the TMB-HRP-MRS agar. The antagonistic effects of the isolates on pathogenic microorganisms were determined with a double layer agar plate. Results One hundred and three lactobacilli strains were isolated from 60 samples of vaginal secretion from healthy pregnant women. Among them, 78 strains could produce hydrogen peroxide, in which 68%, 80%, 80%, and 88% had antagonistic effects against Candida albicans CMCC98001, Staphylococcus aureus CMCC26003, Escherichia coli CMCC44113, and Pseudomonas aeruginosa CMCC10110, respectively. Conclusion The recovery of hydrogen peroxide-producing lactobacilli decreases with the increasing pregnant age and time. The most commonly isolated species from vagina of Chinese pregnant women are Lactobacillus acidophilus and Lactobacillus crispatus. Most of L. acidophilus and L. crispatus produce a high H2O2 level.     

Synthesis and antifungal activity of 1-（1H-1,2,4-triazol-l-yl）-2-（2,4-diflu-orophenyl）-3- （4-substituted acyl piperazin-l-yl）-2-propanols

To study the effect of fluconzole derivatives from a side chain containing 4-substituted acyl piperazin-1-yl on antifungal activity. Methods: Fourteen title compounds were synthesized and confirmed by the elementaryanalysis, ^1HNMR and IR spectra. Five deep fungal strains and 3 shallow fungal strains were chosen as the experimental strains. Minimum inhibitory, concentrations(MICs) of all title coropounds were determined by the method recommended by the National Committee for Clinical Laboratory Standm‘ds (NCCLS) using RPMI 1640 test medium. Results: Among the 14 title compounds, 12 were first reported. The results of preliminary antifungal test showed that all the title compounds exhibited potent antifungal activities to a certain extent. The activity of 4 compounds were more than 4 times as high as that off luconazole and equal to that of ketoconazole against Candida albicans in vitro(MIC80 value≤0.125μg/ml). Conclusion:Introduction of a side chain containing 4-substituted acyl piperazin-1-yl into the main part of fluconazole has important influence on antifungal activities of title compounds.     

Antimicrobial Profile of Lactic Acid Bacteria Isolated from Vegetables and Indigenous Fermented Foods of India against Clinical Pathogens Using Microdilution Method

In dairy and food industries lactic acid bacteria(LAB)have been used in form of starter culture that plays vital role in fermentation;as flavouring and texturizing or as preservative agents.There is increasing evidence that lactobacilli which inhabit the gastrointestinal tract develop antimicrobial activities and participatein the host’s defence system[1].During fermentation,most of the LAB produces a number of different compounds like organic acids,hydrogen peroxide,diacetyl,acetaldehyde,carbon dioxide,polysaccharides,and proteinaceous compounds called bacteriocins or bacteriocinogenic peptides[2-3].Such metabolites exert antimicrobial activity and are     

Clinical Evaluation of ERCP and Naobiliary Drainage for Biliary Fungal Infection——A Report of Five Cases of Severe Combined Bacterial and Fungal Infection of Biliary Tract

Summary： This study studied the use of ERCP and nasobiliary tube in the diagnosis of fungal infection of hiliary tract and the efficacy of combined use of local administration via nasohiliary tube and intravenous antifungal treatment for severe biliary tract fungal infection. 5 patients in our series, with age ranging from 47 to 68 y （mean 55.8）, were diagnosed as having mixed bacterial and fungal infection of hiliary tract as confirmed by smear or/and culture of bile obtained by ERCP and nasohiliary drainage. Besides routine anti-bacteria therapy, all patients received local application of flu- conazole through nasohiliary tube and intravenous administration of fluconazole or itraconazole in terms of the results of in vitro sensitivity test. The mean duration of intravenous fluconazole or itraconazole was 30 days （24-40 days）, and that of local application of fluconazole through nasobiliary drainage tube was 19 days （8 24 days）. During a follow up period of 3-42 months, all patient＇s fungal infection of biliary tract was cured. It is concluded that on the basis of typical clinical features of biliary tract infection, fungal detection of smear/culture of bile obtained by ERCP was the key for the diagnosis of fungal infection of biliary tract. Local application antifungal drug combined with intravenous anti-fungal drugs might be an effective and safe treatment for fungal infection of biliary tract.     

Evaluation of the disc diffusion method with a comparison study for fluconazole susceptibility of Candida strains

Objective To performance susceptibility testing of antifungal agents. Due to the increasing number of resistant strains, susceptibility testing of antifungal agents is gaining importance.Methods We compared the results of standard macrotube dilution reference method with two different microdilution methods, as well as the disc diffusion method in order to test the susceptibility of 150 Candida strains to fluconazole.Results Overall correlation between microdilution and macrodilution methods was 86%. It was 91% between the Minimal Inhibitory Concentrations obtained from macrodUution and disc diffusion zone diameters.Conclusion The disc diffusion test was evaluated as a low-cost, reproducible, and efficient way of assessing the in vitro susceptibility of Candida strains to fluconazole.     

Biliary germiculture and antibiotic susceptibility test in biliary tract infection patients and the selection of antibiotics

Objective To survey the biliary pathogenic bacteria and their resistance to antibiotics in biliary tract infection patients. Methods The data of 326 strains of pathogen isolated from the positive bile culture during January 2004 to December 2005 and their sensitivity to antibiotics were statistically analyzed by WHONET 5.3. Results A total of 326 strains were identified including 176 Gram negative strains （54%, 176/326）,     

Evaluation of the Potential Inhibitory Activity of A Combination of L. Acidophilus, L. Rhamnosus and L. Sporogenes on Helicobacter Pylori: A Randomized Double-Blind Placebo-Controlled Clinical Trial

Objective: To investigate whether three strains of probiotics, L. acidophilus, L. rhamnosus, and L. sporogenes, had significant inhibitive effects on Helicobacter pylori(H. pylori). Methods: This is a 4-week, randomly assigned, parallel-group, doubled-blind, and placebo-controlled study. Fifty patients with a positive H. pylori infection urea breath test(△UBT) result 10% and without ulcer symptoms were randomized into a treatment group and a placebo group by a computer generated allocation sheet with 1:1. These subjects took one capsule of probiotics or placebo twice daily. The primary measurement was the change in △UBT values. Results: The △UBT values during the 4-week treatment period and the 2-week follow-up period were not significantly different between the treatment group and the placebo group, indicating that the inhibitive effects on H. pylori were comparable between both groups. The monocyte count(%) was 5.77±1.11 in the treatment group versus 5.09±1.12 in the placebo group(P=0.044), and the basophile count was 0.55±0.32 in the treatment group versus 0.36±0.23 in the placebo group(P=0.024) at week 2 of the treatment period, both of which reached statistical significance. The monocyte count was 5.75±1.26 in the treatment group and 4.72±0.99 in the placebo group at the end of the follow-up period(P=0.003). Conclusions: There was no significant inhibitive effects of the three probiotic strains(L. acidophilus, L. rhamnosus, and L. sporogenes) on H. pylori. Probiotics can not play the same role as antibiotics in the eradication of H. pylori, the role of probiotics is likely to be important as adjuvant to the triple or quadruple therapy for H. pylori, especially in resistance cases.     

In Vitro and In Vivo Antibacterial Activity of Patchouli Alcohol from Pogostemon cablin

Objective:To investigate the antibacterial activity of patchouli alcohol(PA)against 127 bacteria strains,including the common bacteria and drug-resistant bacteria strains both in the in vitro and in vivo tests.Methods:For the in vitro trial,the antibacterial property of PA against 107 Gram-positive and 20 Gram-negative bacteria strains was screened by agar double dilution method.For the in vivo trial,specific pathogen free Kunming strain of both male and female white mice,were used to test the protective ability of PA after being injected with the median lethal dose of the tested strains.Results:PA possessed antibacterial activity against all the tested 127 strains.In the in vitro test,PA could inhibit both Gram-negative bacteria(25-768μg/m L)and Gram-positive bacteria(1.5-200μg/m L).Particularly,PA was active against some drug-resistant bacteria like methicillin-resistant Staphylococcus aureus(MRSA).PA also exhibited in vivo anti-MRSA activity in mice via intraperitoneal injection.PA could protect mice entirely infected with MRSA at 100 and 200 mg/kg,while 80% mice injected with MRSA could be protected at a low dose of 50 mg/kg.Conclusion:PA might be a potential antibacterial drug from natural sources and might be worthy to explore its mechanism and application in further study.     

COAGGREGATION AND COAGGREGATION INHIBITION BETWEEN PERIO-PATHOGENIC AND CARIOGENIC BACTERIA

Objective To screen the coaggregating pairs between perio-pathogenic and cariogenic bacteria and to investigate the susceptibility of these coaggregations to inhibitors. Methods 4 strains of perio-pathogenic bacteria, Fusobacterium nuleatum （Fn） ATCC 10953, Actinobacilllus actinomycetem comitans （Aa） Y4, Porphyromonas gingivalis （Pg） ATCC 33277,Prevotella intermedia （Pi） ATCC 25261 and 4 strains of cariogenic bacteria, Streptococcus mutans （Sm） lngbritt, Streptococcus sanguis （Ss） 34, Actinomyces viscosus （ Av） 19246 and Lactobacillus acidophilus （La） ATCC 4356 were used to determine the coaggregating degrees of various combinations of the above bacteria by a visual assay and a turbidimetric assay. Then more than ＋ 2 （ or 20% ） coaggregation degrees＇ pairs were used to investigate the inhibitory effect of lactose and arginine and to identify the minimum of their coaggregation-inhibitory concentration. Results The coaggregation degrees of Fn-Av, Pg-Av, Fn-Sm, Fn-Ss, Fn-La and Pg-Ss pairs were higher than ＋ 2 （ 20% ）. 3.0 - 6.0mmol/L of arginine were considerably effective to the above pairs except Fn-Av pair and the disaggregation degrees were 49% - 92%. The maximum of their disaggregation degree to Fn-Av pair was just 18%. 120 - 300mmol/L of lactose were significantly effective to Pg-Ss pair, the disaggregation degrees were 57% - 91%. They partially inhibited Pg-Av pair and were almost ineffective to FnG^＋pairs. Conclusion The coaggregations between perio-pathogenic and cariogenic bacteria are highly specific. Most of them are relatively sensitive to arginine.     

Antifungal susceptibility analysis of berberine, baicalin, eugenol and curcumin on Candida albicans

Objective： To analyze the antifungal effects of Chinese herb monomers, i.e. berberine, baicalin, eugenol and curcumin, on Candida albicans. Methods： After Candida albicans strain Y01-09 was incubated for 48 h in YEPD broth which contained different concentrations of Chinese herb components, the cell cycle, fluorescent intensity and the size of cell volume were detected by flow cytometry. Results： The 4 Chinese herb monomers could affect the cell cycle of Candida albicans in different ranges. The ratio of cells in S-G2-M period decreased as the agents concentration increased, indicating that the cell division was inhibited. The fluorescent intensity of Candida albicans cells became weaker after being incubated, which reflected the loss of DNA fragments. The higher the concentration was, the weaker the fluorescent intensity became. The cell size, cell diopter and particle size changed much as the agents concentration increased. Conclusion： Chinese herb monomers play the antifungal role in inhibiting cell division. FCM could be used to determine the susceptibility of antifungal agents.     

12株乳杆菌益生特性的研究

目的:对12株乳杆菌的益生特性进行研究. 方法:采用活菌计数法对12株乳杆菌的耐酸、耐胆盐特性进行筛选,采用琼脂打孔扩散法检测其抑菌活性,采用细胞培养革兰染色计数法检测其黏附性能. 结果:12株乳杆菌对pH 2.5的强酸和0.3%高胆盐环境均有一定的耐受性,对引起肠道感染的几种病原菌都有不同程度的拮抗,其中约氏乳杆菌JCM1022抑菌谱较广,尤其是对大肠杆菌和肠炎沙门菌有很强的抑菌作用.而罗伊乳杆菌JCM1081对肠上皮样细胞Caco-2具有很强的黏附力. 结论:约氏乳杆菌JCM1022和罗伊乳杆菌JCM1081可作为益生菌候选菌株应用于医药领域.     

几种人源乳酸杆菌对大肠埃希菌的生物拮抗作用

目的：探讨生殖道不同乳酸杆菌菌株对大肠埃希菌的拮抗作用,筛选出作用较强者,用于治疗女性泌尿生殖道感染。方法：采用液体培养法,将乳酸杆菌和大肠埃希菌同时接种于MRS肉汤,在不同时间点取样并进行活埃希菌计数。结果：所选乳酸杆菌都可将大肠埃希菌全部抑杀,但是不同菌株的拮抗效力不同。结论：所选的4株乳酸杆菌对大肠埃希菌的拮抗能力不同,其中I.acidophilus 1和lb．paracasei ssp paracasei 3的抑菌作用发生较快、较强。     

人阴道乳酸杆菌对KM小鼠细胞因子IL-2和IFN-γ浓度影响的研究

目的:探讨四种乳酸杆菌在高、中、低三种浓度的情况下对小鼠细胞因子IL-2和IFN-γ的调节作用并筛选出优良菌株和浓度。方法:用乳酸杆菌专用MRS培养基在厌氧条件下通过几次增菌、分离后,筛选出产过氧化氢的乳酸杆菌,鉴定后选出4株用于本实验。根据四株乳酸杆菌组(L.acidophilus 1 group、L.crispatus group、L.acidophilus 2 group、L.acidophilus 3 group)和三个浓度(1×108、1×109、1×1010),将实验动物随机分为十二组,每组6只。各乳酸杆菌组小鼠腹腔注射相应浓度乳酸杆菌0.2ml/只,另设对照组三组,分别为对照1组(NS1)、对照2组(NS2)、对照3组(NS3),每组6只。各对照组注射生理盐水0.2ml/只,连续10d。处死小鼠用ELISA法检测血清中IL-2、IFN-γ的浓度。结果:①血清IL-2(ng/ml)含量:菌种和剂量总体有差异(P<0.05),四株菌种组与对照组均有差异(P<0.05),Ⅰ组、Ⅱ组、Ⅲ组的IL-2含量高于J组,两者比较有差异(P<0.05),M组高于D组,两者比较有差异(P<0.05);②血清IFN-γ(ng/ml)含量:菌种和剂量总体有差异(P<0.05),四种菌种组与对照组均有差异(P<0.05),Ⅱ组高于Ⅰ组和J组,两者比较有差异(P<0.05),H组高于D组有差异(P<0.05)。结论:乳酸杆菌能调节小鼠T细胞免疫,增强细胞免疫活性,提高血清中IL-2、IFN-γ的浓度;不同菌株和不同浓度各有差异,总体表明:对于细胞免疫的调节作用,高浓度的乳酸杆菌活性大于低浓度乳酸杆菌,Ⅱ型嗜酸乳酸杆菌的活性高于其他三株。     

十种可制作酸奶的乳酸菌体外降解草酸能力评价

目的 评价10种可制作酸奶的乳酸菌在草酸培养液中降解草酸的能力。方法 在5 mmol/L草酸培养液中,分别接种嗜酸乳杆菌、副干酪乳杆菌、屎肠球菌、乳双歧杆菌、青春双歧杆菌、婴儿双歧杆菌、长双歧杆菌、乳酸乳球菌乳脂亚种、保加利亚乳杆菌和嗜热链球菌共10种乳酸菌,培养72 h后检测培养液中草酸浓度和乳酸菌浓度的变化;另设不接种乳酸菌培养的空白对照。结果 经过72 h培养,10种乳酸菌浓度均增加,培养前后差异均有统计学意义（P<0.01）。与空白对照相比,所有接种乳酸菌的培养液草酸浓度均下降,其中嗜酸乳杆菌、乳酸乳球菌乳脂亚种、长双歧杆菌、青春双歧杆菌和乳双歧杆菌培养前后的草酸浓度差异具有统计学意义（P<0.05或P<0.01）;乳双歧杆菌的草酸降解率（29.03％）最高,屎链球菌草酸降解率（0.23％）最低。相关性分析显示,乳酸菌的增殖倍数与草酸降解率无显著相关性（r=0.435 7, P=0.208 2）。结论 10种可制作酸奶的乳酸菌均具有草酸降解能力,草酸降解能力与其在草酸培养液中的增殖能力无相关性。     

人阴道乳酸杆菌的优选和鉴定

目的从本地健康妇女阴道内筛选产H2O2的乳酸杆菌,确定其培养的最适条件,并鉴定出菌种。方法采样后,经过几次增菌培养-平板分离后选出乳酸杆菌,并筛选出产H2O2的菌落进行菌种鉴定。结果分离并鉴定出4种产H2O2的乳酸杆菌,其中L.acidophilusl和Ib.paracaseisspparacasei3产H2O2的能力较强。结论通过试验确定了健康妇女阴道内乳酸杆菌的最佳生长条件,分离并鉴定出4种产H2O2的菌株,为后期实验奠定了基础。     

外阴阴道念珠菌病的临床特征和抗真菌药物敏感性研究

目的探讨外阴阴道念珠菌病（VVC）的临床特征和抗真菌药物敏感性。方法对900例VVC患者采用Soble分类对临床特征进行评分,培养、分离菌株,并对不同菌株进行酮康唑、伊曲康唑、氟康唑、两性霉素B体外药敏试验。结果单纯型VVC的临床特征平均分显著低于非白念珠菌VVC和RVVC（P〈0．05）;689例（76．56％）为白念珠菌,211例（23．44％）为非白念珠菌,两性霉素B的敏感性均为100％,白念珠菌对四种药物的敏感性间有显著性差异（χ2=9．57,P〈0．01）,非白念珠菌对四种药物的敏感性间有显著性差异（χ2=9．21,P〈0．01）。结论白念珠菌是VVC的主要致病菌,念珠菌对唑类药物敏感性有所差异,非白念珠菌的药物敏感性较白念珠菌低,白念珠菌的耐药性不常见。     

嗜酸乳杆菌耗尽培养上清液对兔离体肠平滑肌收缩的影响

目的:观察嗜酸乳杆菌LA14菌株耗尽培养上清液 (spent culture supernatant,SCS)对兔离体肠平滑肌收缩的影响,并初步探讨其作用机制。方法:制备兔离体回肠标本,分别记录肠平滑肌的正常收缩频率和幅度作为给药前对照,然后按累积剂量分别加入嗜酸乳杆菌SCS、活菌菌液、SCS+活菌菌液。每次给药0.3 ml,给药间隔6 min,在每次给药4 min后开始描记收缩曲线,记录时间2 min。观察嗜酸乳杆菌各治疗组对离体肠平滑肌收缩的作用。另取肠段按毛果芸香碱、阿托品或SCS、再毛果芸香碱的顺序给药,观察嗜酸乳杆菌SCS对M型胆碱受体的作用。结果:按累积剂量连续给予SCS或SCS+活菌菌液0.6～1.5 ml后,兔离体肠平滑肌收缩频率明显降低,与给药前比较差异有统计学意义（P<0.05或P<0.01）,其余各点与给药前比较均无统计学差异。SCS、活菌菌液、SCS+活菌菌液在0.3～1.5 ml剂量范围内,仅SCS+活菌菌液在给药达1.5 ml时,兔离体肠平滑肌收缩幅度的降低与给药前比较有统计学差异（P<0.05）,其余各点均无统计学差异。SCS或阿托品可明显对抗毛果芸香碱引起的兔离体肠平滑肌收缩幅度的增加（P<0.05或P<0.01）,SCS还能使兔离体肠平滑肌收缩频率明显减少（P<0.01）。结论:嗜酸乳杆菌SCS可能通过阻断M型胆碱受体抑制兔离体肠平滑肌蠕动。     

52株生殖道念珠菌病病原菌的鉴定及药敏分析

目的分析女性生殖道念珠菌感染状况以及耐药性情况。方法临床标本接种到沙保弱培养基,分离培养出的真菌经革兰染色、芽管形成试验、CHROMagar显色培养进行鉴定,并进行氟康唑、两性霉素B药敏分析。结果120份临床标本分离培养出念珠菌52株。白色念珠菌占69.2%,热带念珠菌占7.7%,克柔念珠菌占5.8%,其他念珠菌占17.3%;分离出的52株念珠菌对氟康唑、两性霉素B敏感率分别为86.5%、94.2%。结论女性生殖道念珠菌感染仍以白色念珠菌为主。念珠菌对氟康唑和两性霉素B的敏感性有差异,应重视念珠菌的培养鉴定和药敏试验,以指导临床合理选择抗真菌药物。     

323株酵母样真菌菌群分布特点及E-Test法药敏分析

目的:探讨2005年度在广东省人民医院检验科细菌室收检各类临床标本中分离到的酵母样真菌的菌群分布特点及其耐药性。方法:用常规培养方法从3140份临床标本中分离出323株酵母样真菌,以美国walk/Away40全自动微生物鉴定仪对所分离的酵母样真菌进行种群鉴定,并用E-Test法做药敏试验。结果:323株真菌中,白色念珠菌、热带念珠菌、光滑念珠菌等分别占63.8%、22.3%和7.4%;对5种常用抗真菌药的耐药率排序依次为:5-氟胞嘧啶(FC)1.9%、两性霉素B(AP)2.2%、酮康唑(KE)5.6%、依曲康唑(IT)10.2%、氟康唑(FL)12.1%。结论:临床上酵母样真菌感染以白色念珠菌和热带念珠菌较常见;真菌耐药菌株的检出率呈上升趋势,临床上必须重视真菌的检出和药敏试验。