健康女性阴道乳杆菌抑菌功能的初步研究

目的:研究健康女性阴道乳杆菌对阴道常见致病菌的抑制作用及其机制。方法:选取北京大学第一医院体检中心18～45岁的育龄期健康女性17例,从其阴道分泌物中分离、纯化乳杆菌并进行鉴定。采用spot on lawn和双层牛津杯法筛选出对粪肠球菌、微小消化链球菌、大肠埃希菌和白假丝酵母菌有抑制作用的阴道乳杆菌菌株。测定乳杆菌的生长曲线及代谢物内的乳酸含量。将乳杆菌与白假丝酵母菌共培养,观察乳杆菌菌体和菌液粗提物在抑制白假丝酵母菌的过程中各自所起的作用。结果:17例健康女性阴道分泌物中分离出23株乳杆菌,其中6株具有抑菌作用,分别是加氏乳杆菌、詹氏乳杆菌、阴道乳杆菌、卷曲乳杆菌、发酵乳杆菌和德氏乳杆菌,其中加氏乳杆菌增殖能力较强,詹氏乳杆菌最弱。6株乳杆菌代谢物内乳酸的含量不同,发酵乳杆菌最高为(2.72±0.035)mg/ml,卷曲乳杆菌和德氏乳杆菌较低,分别为(2.14±0.035)mg/ml和(2.13±0.025)mg/ml。加氏乳杆菌、詹氏乳杆菌、阴道乳杆菌的菌液、菌体及菌液粗提物均可以抑制白假丝酵母菌的生长(P≤0.05);发酵乳杆菌和德氏乳杆菌的菌液和菌体可以抑制白假丝酵母菌的生长(P≤0.05);卷曲乳杆菌对白假丝酵母菌则无抑制作用。结论:健康女性阴道乳杆菌的多个菌株均具有抑制大肠埃希菌、粪肠球菌、微小消化链球菌或白假丝酵母菌的功能。詹氏乳杆菌粗提物中细菌素类物质和过氧化氢的联合作用可以抑制白假丝酵母菌,加氏乳杆菌和阴道乳杆菌主要依靠过氧化氢起作用。乳杆菌产生的乳酸没有发挥抑菌作用。     

细菌性阴道病患者与健康妇女阴道乳杆菌生态学特征的比较

目的:观察健康妇女与细菌性阴道病患者阴道内乳杆菌属内各种乳杆菌在阴道中分布的差异。方法:利用微生态学研究方法,调查45例健康妇女和40例细菌性阴道病患者阴道乳杆菌的分布,鉴定阴道内乳杆菌,比较阴道乳杆菌产生过氧化氢的能力。结果:健康妇女阴道内乳杆菌检出率和平均活菌数量对数值明显高于细菌性阴道病患者(P<0.01),健康妇女阴道内优势乳杆菌是嗜酸乳杆菌(35%,7·68±1.41)、德氏乳杆菌(7·5%,8.06±0.83)和乳酸乳杆菌(7·5%,8.07±1·51),其大多数产生H2O2能力较强;健康妇女阴道内产H2O2乳杆菌的分离率明显高于细菌性阴道病患者(P<0.01)。结论:H2O2阳性的乳杆菌是健康妇女阴道乳杆菌的重要成员,在防治细菌性阴道病发生中起重要作用。     

苦参总碱对阴道常见乳杆菌增殖影响的体外研究

目的探讨苦参总碱对6种阴道常见乳杆菌体外增殖的影响。方法 2018年6月至2019年3月于清华大学附属北京清华长庚医院,将从中国育龄期健康女性阴道中分离、纯化的6种不同阴道乳杆菌进行体外培养,通过琼脂稀释法观察不同浓度的苦参凝胶主要有效成分苦参总碱对各种乳杆菌临床分离株体外增殖的影响。结果苦参总碱对鼠李糖乳杆菌的最低抑菌浓度(MIC)为40 mg/m L;对卷曲乳杆菌、詹氏乳杆菌、加氏乳杆菌、罗伊氏乳杆菌的MIC为20 mg/m L;对阴道乳杆菌的MIC则为10 mg/m L。当苦参总碱浓度低于MIC时,对加氏乳杆菌和罗伊氏乳杆菌具有一定促进增殖的作用,而对卷曲乳杆菌和阴道乳杆菌的增殖无影响。结论临床给药剂量下,苦参凝胶不会抑制阴道常见乳杆菌生长,有利于恢复患者阴道微生态平衡。     

乳杆菌功能改变与外阴阴道假丝酵母菌病

目的研究VVC患者阴道乳杆菌菌种构成与功能特点。方法从VVC患者和健康女性阴道分泌物中分离乳杆菌并鉴定。测定乳杆菌抑制白假丝酵母菌的能力。结果对照组女性阴道分泌物分离出64株乳杆菌,81.3%能抑制白假丝酵母菌增殖,VVC组分离出51株乳杆菌,70.6%能抑制白假丝酵母菌增殖。对照组常见菌种为卷曲乳杆菌、加氏乳杆菌和詹氏乳杆菌,VVC组常见菌种为阴道乳杆菌、发酵乳杆菌和德氏乳杆菌。对照组所有患者均能分离到具有抑制白假丝酵母菌功能的乳杆菌,VVC组仅42%的患者能分离到具有抑制白假丝酵母菌作用的乳杆菌。结论 VVC患者阴道乳杆菌菌种构成与健康女性存在差别。VVC组患者阴道乳杆菌抑制白假丝酵母菌能力较对照组弱。     

需氧菌性阴道炎的诊治进展

需氧菌性阴道炎(aerobic vaginitis,AV)是近年来认识到的一种阴道感染,主要由需氧菌感染引起.AV由Donders等[1]于2002年正式报道,病因及发病机制目前不清楚.正常阴道内以产过氧化氢的乳杆菌占优势.AV时,阴道内能产过氧化氢的乳杆菌减少或缺失,其他细菌主要为需氧菌,如B族链球菌、葡萄球菌、大肠埃希菌及肠球菌等需氧菌增多,并产生阴道黏膜炎性改变.     

细菌性阴道病发病机制及诊治进展

细菌性阴道病（bacterial vaginosis,BV）是育龄妇女常见阴道感染性疾病．可引起多种并发症。BV是多种致病菌作用的结果,阴道内动弯杆菌、普雷沃菌、类杆菌等厌氧菌及加德纳菌、人型支原体大量繁殖,产过氧化氢的乳杆菌减少。诊断标准包括：常用Amsel标准、实验室采用的Nugent革兰染色法及BV快速诊断酶盒和BVBlue检测法等。BV治疗主要有抗厌氧菌药物甲硝唑、克林霉素。复发性BV指BV在1年内反复发作3次或以上,其高复发率是治疗的难点。     

鼠李糖乳杆菌GR-1和罗伊氏乳杆菌RC-14防治阴道炎的研究进展

健康女性阴道内存在以乳杆菌等优势菌为主的微生物菌群,健康的微生物菌群能够维持阴道的酸性环境,增强局部免疫屏障功能,抵御致病菌的侵袭,并减少女性阴道感染的风险。抗生素和抗真菌类药物在杀灭阴道致病菌的同时也抑制了阴道正常有益菌的生长,使阴道菌群失调,导致致病菌占优势,从而出现复发率高、耐药性以及继发感染等问题。口服或者阴道内使用乳杆菌活菌制剂能有效防治女性阴道炎的发生,减少疾病复发。从体内和体外研究两方面,综述鼠李糖乳杆菌（GR-1）和罗伊氏乳杆菌（RC-14）菌株在维持阴道正常菌群平衡、预防和治疗阴道炎中的作用及其研究进展。     

健康妇女和外阴阴道假丝酵母菌病患者阴道乳杆菌对阴道假丝酵母菌抑制作用的实验研究

目的:探讨正常健康和外阴阴道假丝酵母菌病(VVC)妇女阴道分泌物中分离的乳杆菌对假丝酵母菌生长的抑制。方法:健康正常(正常组)和VVC初发(VVC组)妇女各30名,通过定量采集阴道分泌物培养乳杆菌,观察阴道乳杆菌数量及菌种分布情况,并将获得的乳杆菌及VVC患者阴道分泌物中假丝酵母菌分离纯化及鉴定后,采用牛津杯法观察乳杆菌对假丝酵母菌生长的抑制作用,同时比较VVC组和正常组乳杆菌产H2O2能力的差异。结果:①乳杆菌检出率及菌种分布组间无统计学差异(P>0.05);②VVC组阴道乳杆菌数量的平均对数值与正常组相比显著减少(P<0.01);③阴道内乳杆菌BV试剂盒检测为阴性者,正常组显著高于VVC组(P<0.01);④正常组阴道分离的乳杆菌与VVC组比较,对白色和热带假丝酵母菌的生长抑制有统计学差异(P<0.05),对克柔假丝酵母菌的抑制无统计学差异(P>0.05);正常组分离的嗜酸乳杆菌与VVC组的比较,对白色和热带假丝酵母菌的生长抑制均有统计学差异(P<0.01,P<0.05),对克柔假丝酵母菌的生长抑制无统计学差异(P>0.05);但是正常组和VVC组分离的詹氏乳杆菌对上述3种假丝酵母菌的生长抑制均无统计学差异(P>0.05)。结论:①正常妇女与VVC患者阴道内乳杆菌检出率、菌种分布相似,均以嗜酸、詹氏乳杆菌为优势菌种;②VVC患者阴道乳杆菌的数量显著减少;③VVC组产生H2O2的能力下降,可能与VVC发生相关;④正常状态下阴道乳杆菌对假丝酵母菌抑制作用优于VVC感染状态,其中以嗜酸乳杆菌的抑菌作用最为明显。     

细菌性阴道病阴道乳酸杆菌的生态学特征

细菌性阴道病（ｂａｃｔｅｒｉａｌｖａｇｉｎｏｓｉｓ，ＢＶ）系阴道内菌群生理生态内环境改变所致，尤与阴道内乳酸杆菌（ｌａｃｔｉｃａｃｉｄｂａｃｉｌ－ｌｕｓ，ＬＡＢ）与加特纳杆菌（Ｇａｒｔｎｅｒ＇ｓｂａｃｉｌ－ｌａｓ，ＧＢ）或混合性厌氧菌群失调有关。阴道...     

萎缩性阴道炎阴道厌氧菌群的研究

目的 观察萎缩性阴道炎患者阴道的厌氧菌分布。方法 收集萎缩性阴道炎病人５０例、绝经前健康妇女和绝经后阴道萎缩无炎症妇女各２０例，测定阴道ｐＨ值，进行阴道厌氧菌培养。结果 萎缩性阴道炎组的阴道ｐＨ值（７．０）明显高于绝经前妇女（４．６），共分离出厌氧菌５６株，分离率由高到低前四位依次是双向拟杆菌７例（１４％）、产生消化链球菌７例（１４％）、消化链球菌５例（１０％）、不解乳真杆菌４例（８％），萎缩性阴     

内蒙古牧区育龄期健康及细菌性阴道病妇女阴道乳杆菌种的鉴定与分析

目的 对内蒙古牧区育龄期健康妇女及细菌性阴道病(BV)患者的阴道乳杆菌种进行鉴定与分析,为阴道微生态治疗提供依据.方法 2008年6至12月,选取内蒙古自治区3个牧区旗县(正镶白旗、达尔罕茂明安联合旗、百灵庙镇)育龄期蒙古族健康妇女203例、汉族健康妇女74例,蒙古族BV妇女102例,采用改良乳杆菌培养基(MRS)对阴道侧壁分泌物中乳杆菌进行分离、培养,提取细菌总DNA,进行核糖体脱氧核糖核酸(16S rDNA)基因序列鉴定,并采用四甲基联苯胺-辣根过氧化物酶-乳杆菌培养基(TMB-HRP-MRS)对乳杆菌进行产H2O2分析.结果 (1)检出率:203例蒙古族健康妇女乳杆菌检出率为76.8%(156/203),蒙古族BV妇女乳杆菌检出率为21.6%(22/102),两者比较,差异有统计学意义(P＜0.01);汉族健康妇女乳杆菌检出率为82.4%(61/74),与蒙古族健康妇女比较,差异无统计学意义(P＞0.05).(2)阴道乳杆菌鉴定:蒙古族健康妇女阴道分泌物中共检出193株乳杆菌株,共11种,蒙古族BV妇女阴道分泌物中检出22株乳杆菌株,共4种.(3)H2O2阳性乳杆菌的鉴定:对分离出乳杆菌的74例蒙古族健康妇女及22例蒙古族BV妇女进行阴道乳杆菌H2O2实验,其中蒙古族健康妇女阴道H2O2阳性乳杆菌的检出率为75.7%(56/74),蒙古族BV妇女阴道H2O2阳性乳杆菌的检出率为27.3%(6/22),两者比较,差异有统计学意义(P＜0.05).结论 内蒙古牧区育龄期健康妇女阴道乳杆菌检出率与种族无明显相关性;BV妇女的阴道乳杆菌及H2O2阳性乳杆菌检出率均较健康妇女明显降低.

Abstract：

Objective To identify and analyze the species of vaginal lactobacilli between patients with bacterial vaginosis (BV) and healthy women at childbearing age in Inner Mongolia. Methods From Jun. 2008 to Dec. 2008, 203 Mongolian healthy women, 74 Han healthy women and 102 Mongolian patients with BV from 3 pastoral areas were enrolled in this study. Isolation and culture of lactobacilli from vaginal wall were performed by modified culture medium. DNA of lactobacilli were extracted and sequenced. H2O2 were detected by TMB-HRP-MRS. Results(1)The rate of lactobacilli identification were 76.8%(156/203) in Mongolian healthy women and 21.6% (22/102) in Mongolian patients with BV, which reached statistical difference(P＜0.01).Lactobacilli identification in Han healthy women [82.4%(61/74)] did not show significant difference with that of Mongolian healthy women (P＞0.05). (2) The total of 193 strains and 11 species of Lactobacillus were detected in 203 Mongolian healthy women. Meanwhile,22 strains and 4 species of Lactobacillus were found in 102 Mongolian BV cases.(3)The rate of H2O2 generating Lactobacilli was 27.3% (6/22) in Mongolian BV patients and 75.7% (56/74)in Mongolian healthy women, which showed statistical difference(P＜0.05). Conclusions The rate of Lactobacillus was not related with the race of women in pastoral area in Inner Mongolian. The amount of lactobacilli and H2O2 generating Lactobacilli in the vagina of BV patients was remarkably lower than those of healthy women at childbearing age.     

Phage infection in vaginal lactobacilli: an in vitro study

Objective: During bacterial vaginosis, an unexplained decrease of vaginal lactobacilli occurs. To identify whether these lactobacilli could be infected by phages, we isolated phages from vaginal lactobacilli and analyzed their potential virulence in attacking vaginal lactobacilli in vitro.Methods: Vaginal samples were obtained from 39 reproductive-aged women. The selective Rogosa SL agar was used to isolate lactobacilli, from which phages induced by mitomycin C or released spontaneoulsy were analyzed by the agar spot method.Results: Of 20 samples from women with vaginal infections, 12 did not have lactobacilli. From the remaining 8 infection samples and the 19 samples from healthy women, 37 Lactobacillus strains were isolated, from which 7 temperate phages were identified. Upon analysis, all 7 phages infected vaginal lactobacilli from the same and/or different women in vitro. Two phages, Phikc005 and Phikc007, had a broad host range, infecting 7 of 8 species tested. A control intestinal Lactobacillus phage also lysed several vaginal strains. One vaginal phage, Phikc039, was apparently lytic against vaginal lactobacilli from 7 other women. This phage was characterized as follows: plaque morphology, small and clear; burst size, 300 phages per cell; spontaneous induction rate, 1 per 10(6) cells; DNA, double-stranded and linear, 41 kb; and shape, a hexogonal head and a non-contractile tail.Conclusions: Bacteriophages were isolated from vaginal lactobacilli of some women and were shown in vitro to lyse vaginal Lactobacillus strains from the same and/or different women. It was suggested that vaginal lactobacilli might be suppressed by phages.     

Lactobacilli in the female genital tract in relation to other genital microbes and vaginal pH

BACKGROUND: The relationship between lactobacilli and other microbes and the association with vaginal pH in the female genital tract were examined. The study also included evaluation of the possibility of supplying probiotics to the genital tract by using panty liners impregnated with the probiotic strain Lactobacillus plantarum LB931. METHODS: This was a randomized, placebo-controlled, double-blind, multicenter study involving 191 healthy fertile women. Specified microbes were counted and vaginal pH was measured once a month for five consecutive months. RESULTS: Major individual variations in the genital microflora composition and the vaginal pH were found among the women. The number of lactobacilli was significantly related to vaginal pH (p<0.001) and approximately 70% of the women were permanent carriers of individual lactobacilli strains. Women with high numbers of lactobacilli were less prevalent with Group B streptococci than women with low numbers (p=0.036), and these women had a lower mean vaginal pH. The number of lactobacilli also correlated with the prevalence of yeast. LB931 could be found in 86% of the labial samples and 54% of the vaginal samples. CONCLUSIONS: High numbers of lactobacilli may contribute to a low vaginal pH and seem to have a negative influence on Group B streptococci. LB931 could be transferred from the panty liners to both the vagina and the labial fold.     

Could bacterial vaginosis be due to the competitive suppression of lactobacilli by aerobic microorganisms?

Bacterial vaginosis (BV) is characterized by lack of lactobacilli and predominance of anaerobic species. Ciprofloxacin is active against a great number of aerobic bacteria. The effect of ciprofloxacin on the vaginal microbial flora was studied both in healthy subjects and in patients with BV in order to test if BV may be due to competitive suppression of lactobacilli by aerobic species. About half of the patients were relieved of their symptoms. No adverse effects on the microbial profile or colonization by lactobacilli were produced by the drug. Lactobacilli known to be able to produce hydrogen peroxide were found to be significantly more common in healthy women than in women with BV. The species most frequently related to vaginal health were Lactobacillus jensenii and Lactobacillus rogosae.     

Hydrogen peroxide producing lactobacilli in women with vaginal infections

OBJECTIVE: Hydrogen peroxide producing lactobacilli are isolated from the vaginas of a majority of healthy reproductive age women. Their toxic and inhibitory effect against the overgrowth of pathogens in the vagina is documented by in vitro studies. Clinical studies concerning the role of hydrogen peroxide producing lactobacilli in vaginal infections are controversial. STUDY DESIGN: The aim of this study was to isolate lactobacilli from women with major vaginal infections: bacterial vaginosis (BV), vulvovaginal candidiasis (VVC) and trichomoniasis (TV), and from women with normal flora (NF), to test their ability to produce hydrogen peroxide, to quantitate lactobacilli and to evaluate factors related to the colonization of vagina with hydrogen peroxide producing lactobacilli. The study group comprised 108 women: 27 with VVC, 26 with BV, 20 with TV and 35 with NF. Data was analysed with Fisher exact test, chi2-test, and ANOVA. RESULTS: Lactobacilli were isolated from 81.5% of women with VVC, 58% of women with BV (p=0019), 55% of women with TV (p=0.015) and from 86% of women with NF. The majority of isolates of lactobacilli from women with VVC, TV and NF produced hydrogen peroxide (77%, 63.5% and 80%, respectively), compared with women with BV where only 26.5% of isolates produced hydrogen peroxide (p=0.01). Quantitative analysis showed that in an overwhelming majority of women colonized with lactobacilli, their count was >10(7) CFU/ml. Upon evaluation of behavioural and demographic characteristics, we found that smokers had a lower incidence of isolation of hydrogen peroxide producing lactobacilli (p=0.018). Our results showed that women with BV lack lactobacilli, especially hydrogen peroxide producing ones. Women with TV had a lower rate of lactobacilli. In women with VVC, there was neither significant difference in the isolation of lactobacilli, nor in their ability to produce hydrogen peroxide, compared to women with NF. CONCLUSION: Our results point out that hydrogen peroxide producing lactobacilli could protect against the development of BV, but not against VVC and TV.     

The relationship of hydrogen peroxide-producing lactobacilli to bacterial vaginosis and genital microflora in pregnant women.

Lactobacilli provide an important microbial defense against genital colonization by pathogens. The role of hydrogen peroxide (H2O2) in the control of genital microflora was explored in a cross-sectional study of 275 women in the second trimester of pregnancy. Vaginal cultures were obtained for detection of H2O2-positive and H2O2-negative lactobacilli and other members of the genital microflora. Compared with women with H2O2-negative lactobacilli, women colonized by H2O2-positive lactobacilli were less likely to have bacterial vaginosis, symptomatic candidiasis, and vaginal colonization by Gardnerella vaginalis, Bacteroides, Peptostreptococcus, Mycoplasma hominis, Ureaplasma urealyticum, and viridans streptococci (P less than or equal to .05 for each comparison). In addition to the above organisms, women without vaginal lactobacilli were more likely than those women with H2O2-positive lactobacilli to have Chlamydia trachomatis, and less likely to be colonized by Enterococcus or coagulase-negative staphylococci (P less than .05 for each comparison). Vaginal colonization by group B streptococci or Escherichia coli was not related to the presence of H2O2-positive lactobacilli. These data suggest that the presence of H2O2-positive lactobacilli in the vagina is inversely correlated with infection by some genital pathogens in pregnant women.     

Long, uniform Lactobacilli (Döderlein's Bacteria): a new risk factor for postoperative infection after first-trimester abortion

Objective: The production of hydrogen peroxide (H₂O₂) from different strains of lactobacilli in the vagina has been proposed to play one of the most important protective roles in the vaginal defense system. New data have, however, suggested that Döderlein''s bacteria, with the morphological appearance of long lactobacilli, have a low production of H₂O₂ . The purpose of the present study was to correlate the morphology of lactobacilli with the incidence of infection following legal abortion.Methods: Seven hundred sixty-nine women with lactobacilli but without Chlamydia trachomatis or bacterial vaginosis in their vaginal wet smears who were to undergo legal abortions were divided into 6 different groups according to the morphological appearance of the lactobacilli. The postoperative infection rates in these different groups were compared. A phenotypic classification of some of the lactobacilli was performed.Results: The presence of Döderlein''s bacteria compared with a mixed flora of lactobacilli increased the risk of postoperative infection 3 times [relative risk (RR) = 3.0; 95% confidence interval (CI), 1.5-6.3]. After a logistic regression analysis, the only independent risk factors were the presence of Döderlein''s bacteria and earlier gestational age.Conclusions: We found that the lactobacilli regarded as commensal organisms and “normal, healthy lactobacilli” in the vagina were present in only 18% of these women and that their presence might be hazardous. Therefore, we must reconsider our concept of the “normal” lactobacilli in the vaginal wet smears of healthy women.     

Bacterial flora of the cervix in women using different methods of contraception

Bacteriologic culture samples were taken from the cervix in three groups of 10 healthy, sexually active women using barrier contraception, oral contraceptives, or a levonorgestrel-releasing intrauterine contraceptive device. Culture samples for Candida albicans and Trichomonas vaginalis were taken, a cytologic vaginal smear was obtained, and an amine sniff test was performed; these were in addition to a routine gynecologic examination. Multiple bacteria were isolated from the cervix in women using oral contraceptives or an intrauterine contraceptive device, whereas lactobacilli alone dominated the flora of women using barrier contraception. Significantly more anaerobic bacteria were isolated from the cervix in oral contraceptive and intrauterine contraceptive device users when compared with the barrier method users. Symptoms and findings evident of anaerobic vaginosis were associated with the occurrence of anaerobic bacteria in the cervix of three patients using the intrauterine contraceptive device. The results showed that the cervical bacterial flora in sexually active healthy women is rich in anaerobes that can be regarded as a normal finding in women using oral contraceptives or intrauterine contraceptive devices. Barrier contraception with a condom prevents this anaerobic shift and maintains a lactobacilli-dominated flora in the cervix.     

Influence of phenoxymethylpenicillin on the vaginal ecosystem.

The effects of phenoxymethylpenicillin on the bacterial flora of the vagina were investigated in 6 women who fulfilled the criteria of 'normality': predominance of lactobacilli, and absence of Enterobacteriaceae and fungi. Fourteen healthy women were enrolled and followed longitudinally; 8 of these were subsequently excluded because of divergence from normality. Following penicillin treatment, Gram-negative rods appeared in 4 of the remaining 6 women, and in 1 subject heavy growth of Candida albicans resulted. In 5 of the women there was no change in the lactobacillus count and a rise in pH was found in 4 subjects. In 1 woman, lactobacilli disappeared completely and were replaced by Escherichia coli. Concomitantly, high concentrations of endotoxins were found in the vaginal fluid. Treatment with penicillin may interfere with the normal vaginal bacterial flora and provide an environment favorable for Enterobacteriaceae and fungi.