Antimicrobial activity and protective properties of vaginal lactobacilli from healthy Bulgarian women

The role of vaginal Lactobacillus as an efficient barrier against invading pathogens is of considerable interest. The purpose of the present study was to assess in vitro the ability of 20 recently identified vaginal lactobacilli to protect the vagina. In order to evaluate their significance, the antimicrobial, hemagglutination (HA) and aggregation (Agg) activities, as well as acid and hydrogen peroxide (H(2)O(2)) production, were estimated. The cell-free cultures of eight strains showed a stable antimicrobial activity after elimination of the putative effects of lactic acid and H(2)O(2). Three of the isolated vaginal lactobacilli expressed a broad spectrum of anti-bacterial activity including Gram-negative pathogens. Strains with anti-Gardnerella and anti-herpes simplex virus type 2 activities were found. All tested isolates were H(2)O(2) producers, actively acidifying the growth media to pH 3.92+/-0.04, which is presumed to neutralize sexually transmitted infection pathogens. The major part (75%) expressed an HA activity and different Agg phenotypes, estimated as important properties in the competition with invading pathogens and in host defense. These results are encouraging and prompt further research of the characterized active strains and their possible application in prophylaxis of vaginal disorders.     

门诊妇女阴道乳酸菌对临床常用抗生素耐药性的初步研究

目的研究临床常用抗生素类药物对妇女生殖道正常菌群乳酸菌的影响,为临床医生选用抗生素治疗妇科疾病提供初步参考,并为微生态学治疗和预防相关疾病提供理论依据。方法采用K-B法（纸片扩散法）,以临床采集到的12株阴道乳酸菌为试验菌,研究其对临床常用9大类13种抗生素的耐药性。结果在所测试的抗生索中,试验菌对甲硝唑形成的耐药环直径最小,对头孢噻肟形成的耐药直径最大。除甲硝唑外不同菌株对不同抗生索形成的耐药环直径差异较大。结论在该试验的条件下,乳酸菌对甲硝唑耐药性最强,可作为治疗阴道感染的首选药物;乳酸菌对头孢噻肟最敏感,因而在治疗阴道感染时,应尽量避免使用该药物。     

Susceptibility of vaginal bacteria to metronidazole and tinidazole

In vitro antimicrobial susceptibility testing was performed on 470 vaginal isolates from women with bacterial vaginosis and three species of Lactobacillus, to metronidazole and tinidazole using the agar dilution method. There was no significant difference observed in the inhibitory activity of either drug to any of the isolates tested.     

乳杆菌属细菌对宫颈上皮内瘤变患者阴道菌群的影响

目的 分析乳杆菌属细菌对宫颈上皮内瘤变(CIN)患者阴道菌群的影响,为该类患者的治疗提供参考.方法 选择2019年1月至2020年1月我院收治的CIN患者137例,根据组织学病理结果分为CIN Ⅰ组(84例)和CIN Ⅱ、Ⅲ组(53例).选择同期100例宫颈检查正常者为对照组.比较各组对象阴道菌群数量.根据患者阴道内乳...     

Rapid identification of the three major species of dairy obligate heterofermenters Lactobacillus brevis, Lactobacillus fermentum and Lactobacillus parabuchneri by species-specific duplex PCR

In this study, the biodiversity of 154 strains of lactic acid bacteria, including 112 dairy product isolates presumptively identified as obligately heterofermentative lactobacilli (OHL) by classical microbiological tests, as well as 23 OHL-type strains, was investigated by PCR-based methods and gene sequencing. Using these techniques, 51% of the cheese isolates were actually identified as OHL. The non-OHL isolates were identified to the Leuconostoc, Lactobacillus, Weisella, Pediococcus or Streptococcus genera. Among the OHL cheese isolates, five species were directly identified including three of the most frequently isolated species -Lactobacillus fermentum, Lactobacillus parabuchneri and Lactobacillus brevis- and two rarely isolated species - Lactobacillus diolivorans and Lactobacillus reuteri. A sixth group made up of two dairy isolates was also identified and according to 16S rRNA gene and intergenic spacer region (ISR) sequencing data corresponded to Lactobacillus sp. and may constitute a new species. Species-specific primers were designed for the rapid and reliable detection of the three most frequently isolated species by species-specific duplex PCR.     

小羽贯众提取物抗菌活性的初步研究

目的：比较小羽贯众根状茎和叶提取物体外抑菌活性。方法：牛津杯法和96孔板法,通过测定抑菌圈大小、最小抑菌浓度（MIC）和最小杀菌浓度（MBC）,比较小羽贯众根状茎和叶提取物分别对10种常见细菌的抑制作用,以及不同产地抑菌效果的比较。结果：小羽贯众的根状茎和叶的提取物对10种常见的细菌均表现出较强的抑菌活性,肺炎克雷伯氏菌对提取物最敏感。结论：小羽贯众提取物作为一种天然的抗菌物质,具有广谱抗菌效果,有较高的应用潜力和开发前景。     

乳杆菌制剂治疗阴道混合感染的临床疗效观察

目的探讨乳杆菌活菌制剂联合不同药物治疗阴道混合感染的效果。方法 2011年2月至2011年5月在厦门市妇幼保健院确诊为阴道混合感染的117例患者,其中,VVC+AV感染51例(43.59%)、BV+AV感染49例(41.88%)、TV+AV感染5例(4.27%)、VVC+BV+AV感染5例(4.27%)、TV+BV+AV感染5例(4.27%)、TV+BV感染3例(2.53%)。患者在应用相应抗菌制剂的同时联合应用乳杆菌活菌胶囊(定君生),并于下次月经干净后再次应用乳杆菌活菌胶囊1疗程,巩固治疗。观察首次及巩固治疗后的有效率和复发情况。结果 117例患者首次治疗后痊愈82例,显效21例,有效率为87.18%;巩固治疗后痊愈94例,显效19例,有效率为96.58%。二者差异有统计学意义(P<0.05)。结论乳杆菌活菌胶囊(定君生)联合相应药物治疗阴道混合感染临床疗效好,无严重并发症,值得临床推广应用。     

Growth inhibition of Staphylococcus aureus by H2O2-producing Lactobacillus paracasei subsp. paracasei isolated from the human vagina

H2O2 production by certain Lactobacillus strains is one of the mechanisms that helps to regulate the vaginal ecosystem. This paper describes the kinetics of H2O2 production by two different strains of Lactobacillus paracasei subsp. paracasei under different culture conditions and the effect of this metabolite on the growth of Staphylococcus aureus. L. paracasei F2 produced 2.72 mmol 1-1 H2O2 while L. paracasei F28 produced 1.84 mmol l(-1), both in agitated cultures. Although L. paracasei F2 produced a higher H2O2 concentration than L. paracasei F28, H2O2 production per number of live bacterial cells was 10-fold higher for F28. The latter also showed a faster decrease in viability during the stationary phase. There were no detectable levels of H2O2 in cultures without agitation. H2O2-producing lactobacilli inhibited growth of S. aureus in a plaque assay and in mixed cultures, depending on the initial inoculum of the pathogen.     

小羽贯众提取物抗菌活性的初步研究

目的:比较小羽贯众根状茎和叶提取物体外抑菌活性。方法:牛津杯法和96孔板法,通过测定抑菌圈大小、最小抑菌浓度(MIC)和最小杀菌浓度(MBC),比较小羽贯众根状茎和叶提取物分别对10种常见细菌的抑制作用,以及不同产地抑菌效果的比较。结果:小羽贯众的根状茎和叶的提取物对10种常见的细菌均表现出较强的抑菌活性,肺炎克雷伯氏菌对提取物最敏感。结论:小羽贯众提取物作为一种天然的抗菌物质,具有广谱抗菌效果,有较高的应用潜力和开发前景。     

Antimicrobial susceptibility of anaerobic bacteria in Bulgaria

ObjectivesThe antimicrobial susceptibility of anaerobic bacteria isolated from clinical specimens in the referent for Bulgaria anaerobic laboratory was studied in a period of 25 years/1983–2007/.MethodsNCCLS – recommended agar dilution methods were used. β-lactamase activity was determined with nitrocefin discs.ResultsThe 29 antimicrobial agents included in the study were divided according to their in vitro activity against the anaerobic isolates into 4 main groups for guiding empirical treatment: 1st group of metronidazole, chloramphenicol, meropenem, imipenem and combinations of β-lactam antibiotics with sulbactam – with high activity and drugs of choice for treatment; 2nd group – clindamycin, cefoxitin, carbenicillin/and azlocillin, piperacillin/ – with a good activity and low percent of resistant strains; 3rd group – of tetracycline and erythromycin with higher percent of resistant strains including the new macrolides as josamycin, clarithromycin, roxithromycin and azithromycin; 4th group – penicillins/ampicillin, amoxicillin, penicillin/and cephalosporins/cefamandole, cefazolin, cefotaxime and cefoperazone/ – not suitable for treatment of infections including Bacteroides fragilis group strains, with a very high percent of resistant strains, probably due to β-lactamase activity in most of the strains.ConclusionA continued updating and a follow-up in the changes of antibiotic susceptibility are necessary in every country as resistance patterns vary not only between geographical regions but also even among medical centers and hospitals which may be connected with differences in antibiotic usage in man and animals.     

阴道乳杆菌与宫颈癌相关性研究进展

人类阴道微生物群是由200多种细菌组成的复杂系统,其组成受到基因、民族背景、行为以及环境因素等的影响。其中乳杆菌是女性阴道内的优势菌,在健康女性的阴道内主要有L. iners、L. crispatus、L. gasseri和L. jensenii这4种乳杆菌。阴道内乳杆菌主要通过代谢阴道上皮细胞内的糖原产生乳酸,维持阴道正常的酸性环境。此外,阴道乳杆菌还可通过产生过氧化氢(H2O2)和细菌素等物质发挥抵御致病微生物入侵的作用。近年来的研究显示女性阴道HPV感染与阴道微生态失衡尤其是乳杆菌含量的减低有关。本文将围绕阴道内乳杆菌的生理作用及其与宫颈癌发病的相关性作一综述。     

温郁金内生拮抗细菌B-11的分离及其抑菌活性

【背景】植物内生菌广泛分布在自然界中,具有巨大的潜在开发与应用价值。【目的】对温郁金内生细菌进行分离鉴定,并从中筛选出具有生防潜能的菌株。【方法】采用常规组织分离法对温郁金根茎内生细菌进行分离并利用16S rRNA基因序列分析进行初步鉴定;采用平板对峙法以铁皮石斛炭疽病菌(Colletotrichum gloeosporioides)为供试菌株对温郁金内生细菌进行拮抗菌的筛选;通过形态学鉴定、生理生化鉴定以及16S rRNA基因序列分析,确定拮抗菌株B-11的分类地位;以6个不同属的植物病原真菌为供试菌株对拮抗菌株B-11的抑菌谱进行测定,并研究其对病原菌菌丝的影响;利用特异性平板和MALDI-TOF-MS检测技术对拮抗菌株B-11产生的抑菌物质进行检测;采用离体叶片接种法研究拮抗菌株B-11对铁皮石斛炭疽病菌的防治效果。【结果】从温郁金根茎中分离得到25株内生细菌菌株,这些细菌菌株分属于12个属,其中芽孢杆菌属为优势菌属,占分离菌株的28%;通过初筛获得8株对C.gloeosporioides有抑菌活性的菌株,其中菌株B-11的抑菌活性最强,经鉴定该菌为贝莱斯芽孢杆菌(Baclliusvelezensis);抑菌谱测定发现菌株B-11能够对供试的属于不同属的6种植物病原真菌的菌丝生长具有抑制作用,显微观察发现经对峙培养6 d后这6种病原菌的菌丝体出现畸形膨大、分枝增多等现象;特异性平板检测结果表明拮抗菌株B-11能够产生蛋白酶、β-葡聚糖酶和嗜铁素,但不产生几丁质酶;MALDI-TOF-MS检测结果表明拮抗菌株B-11能够产生伊枯草素、丰源素和表面活性素3种脂肽类抗生素,其中伊枯草素的产量最高;离体叶片接种实验表明,拮抗菌B-11的离心去菌发酵液对铁皮石斛炭疽病的防治效率可达64%。【结论】温郁金根茎含有丰富的内生细菌资源,其内生细菌菌株B-11有潜力作为开发抗真菌代谢物和新药物的重要微生物资源。     

乳酸菌阴道胶囊辅助治疗对未足月胎膜早破的疗效及对其阴道微生态的影响

目的 观察乳酸菌阴道胶囊辅助治疗对未足月胎膜早破的治疗效果,并探讨其对阴道微生态的影响。 方法 86例未足月胎膜早破患者采用随机数表分为常规组与研究组,各43例。常规组予以常规治疗,研究组予以乳酸菌阴道胶囊联合常规治疗。比较治疗后两组阴道微生态、胎儿宫内感染率和分娩方式、母体和胎儿不良妊娠结局发生情况。 结果 治疗后研究组Chao1指数和Shannon指数均高于常规组(均P结论 乳酸菌阴道胶囊辅助治疗未足月胎膜早破相较于常规治疗可改善阴道微生态,还可降低胎儿宫内感染率、剖宫产率,减少母体和围产儿不良妊娠结局的发生。     

乳酸菌阴道胶囊对宫颈癌TP化疗患者炎症及阴道菌群的影响

目的 探讨乳酸菌阴道胶囊对中晚期宫颈癌TP化疗患者血清IL 2、IL 10、TGF β1水平及阴道菌群的影响。 方法 选取2018年10月-2019年8月于本院接受TP化疗的83例中晚期宫颈癌患者为研究对象,以随机数字表法分为两组,观察组43例,对照组40例。两组均接受TP化疗,观察组在对照组基础上接受乳酸菌阴道胶囊治疗。比较两组临床疗效,治疗前后血清IL 2、IL 10、TGF β1水平,阴道菌群分布情况,致病微生物阳性检出率和化疗相关不良反应发生情况。 结果 两组临床疗效等级分布差异显著(P0.05)。 结论 乳酸菌阴道胶囊可提高中晚期宫颈癌TP化疗患者临床疗效,调节患者免疫应答水平,有效改善患者阴道菌群平衡,降低患者阴道致病微生物阳性率,安全性高。     

乳杆菌优良菌株的筛选及对小鼠阴道白假丝酵母菌过度增殖干预

目的探讨乳杆菌优良菌株的筛选及其对小鼠阴道白假丝酵母菌过度增殖干预作用。方法对德氏乳杆菌DM8909菌株进行诱变筛选,得到耐抗生素突变株208,以ICR小鼠为实验对象,用甲硝唑进行小鼠阴道菌群脱污染,将白假丝酵母菌接种到小鼠阴道内,构建小鼠阴道白假丝酵母菌过度增殖模型。采用阴道接种德氏乳杆菌DM8909突变株208菌液（2×108CFU/mL）处理,取其阴道冲洗液进行阴道菌群分析,并进行阴道上皮细胞的电镜检查。结果乳杆菌干预后显著清除阴道内的假丝酵母菌,肠杆菌恢复正常水平,与模型组比较,差异有统计学意义（P〈0.05）。结论德氏乳杆菌DM8909抗性菌株208能调整白假丝酵母菌在小鼠阴道内定植,对小鼠阴道上皮细胞有恢复作用。     

Antimicrobial susceptibility of non-O1 Vibrio cholerae isolated from wastewater stabilization ponds in Marrakesh,Morocco

The 365 strains of Vibrio cholerae, isolated in Marrakesh from raw sewage and stabilization pond effluent, were all identified as non-O1 Vibrio cholerae. When tested for their susceptibilities to ampicillin, amoxicillin, cephalothin, streptomycin, novobiocin, chloramphenicol, nalidixic acid and trimethoprim-sulphamethoxazole, 13% of the strains from raw sewage and 20% of those from stabilization pond effluent were found to be resistant to one or more of the antibiotics. There were no significant differences, in terms of drug resistance, between isolates from the new sewage and those from the ponds' effluent.The authors are with the Université Cadi Ayyad, Faculté des Sciences Semialia, Département de Biologie, Laboratoire de Microbiologie, BP S/15, Marrakesh, Morocco     

中国汉族育龄女性阴道内乳杆菌的鉴定与系统发育分析

目的比较基质辅助激光解吸电离飞行时间质谱(MALDITOF MS)与16S rRNA基因测序鉴定女性阴道乳杆菌方法,同时探究不同阴道微生态背景的汉族育龄女性阴道内乳杆菌的种类及其分布情况。方法对260例汉族育龄女性阴道拭子进行阴道微生态分析及乳杆菌的分离培养,根据阴道微生态鉴定结果分为正常组(173例)、中间型细菌性阴道病组(IBV,55例)、细菌性阴道病组(BV,9例)和外阴阴道炎假丝酵母菌病组(VVC,23例),分离乳杆菌并进行纯化培养,纯化的乳杆菌经MALDITOF MS和16S rRNA基因测序鉴定并进行聚类分析。结果2种方法鉴定的一致率为95.3%。260例标本中149例(57.3%)检出了乳杆菌,共发现17种乳杆菌,检出的乳杆菌分布于5个乳杆菌系统发育群,其中分离出较多的乳杆菌为卷曲乳杆菌(47例)、加氏乳杆菌(38例)、阴道乳杆菌(21例)。正常组116例(67.1%)标本分离出乳杆菌,IBV组19例(34.5%),BV组2例(22.2%),VVC组12例(52.2%)。正常育龄女性与患BV女性乳杆菌种类与数量差异有统计学意义(χ²=22.936 0,P<0.000 1),与患VVC女性差异无统计学意义(χ²=1.983 0,P=0.160 0)。与其他组相比,正常组育龄女性中分离出卷曲乳杆菌的比例显著高于BV组和VVC组(χ²=4.548 0,P=0.033 0;χ²=5.605 0,P=0.020 0),VVC组育龄女性分离出加氏乳杆菌的比例显著高于正常组(χ²=5.090 0,P=0.035 0)。结论对比16S rRNA基因测序与MALDITOF MS方法的乳杆菌鉴定结果,除了阴道乳杆菌和人阴道乳杆菌出现不一致的鉴定结果,其余15种乳杆菌鉴定结果一致性均为100%,但基于测序与基于质谱的方法产生的乳杆菌聚类分析结果不完全一致。卷曲乳杆菌可能是反映育龄女性正常阴道微生态的重要标志物,加氏乳杆菌可能与女性VVC的发生有关。     

应用qPCR方法快速定量检测阴道分泌物中乳杆菌的研究

目的建立快速、定量检测女性阴道分泌物中乳杆菌的方法。方法合成针对乳杆菌的特异性引物,建立乳杆菌qPCR标准曲线后,进一步对收集得到的健康女性及细菌性阴道病女性阴道分泌物样品中乳杆菌进行定量检测。结果引物仅扩增出乳杆菌特异性DNA条带,qPCR能够检测到各分泌物样本中乳杆菌的数量。结论建立了一种快速、定量女性阴道分泌物中乳杆菌检测方法。