阴道中益生特性乳杆菌的分离与功能评价

目的 分离健康女性阴道中的乳杆菌并鉴定其益生特性，为开发治疗妇科疾病的复方益生菌制剂提供新型菌株。方法 采集健康女性阴道分泌物并分离筛选乳杆菌，通过16S rDNA序列分析鉴定乳杆菌分离株，并对其产酸性能、产H2O2能力、抑菌能力、产生物膜能力进行检测。结果 从50名健康女性阴道内共分离出179株乳杆菌，其中卷曲乳杆菌101株、詹氏乳杆菌42株、格氏乳杆菌26株、植物乳杆菌5株、唾液乳杆菌3株以及干酪乳杆菌2株。179株乳杆菌中有146株具有产酸能力，发酵液pH值的最低的5株菌分别为卷曲乳杆菌J3、卷曲乳杆菌J8、詹氏乳杆菌J87，植物乳杆菌J75以及格氏乳杆菌J35，其pH分别为4.20、4.23、4.24、4.26及4.36；产H2O2弱阳性菌株有87株、阳性有37株、强阳性有9株，这9株菌分别为卷曲乳杆菌J3、卷曲乳杆菌J8、卷曲乳杆菌J20、詹氏乳杆菌J87，詹氏乳杆菌J90、詹氏乳杆菌J15、格氏乳杆菌J11、植物乳杆菌J75、植物乳杆菌J69以及植物乳杆菌J40；能拮抗大肠埃希菌的菌株有115株、拮抗金黄色葡萄球菌的有84株、拮抗白假丝酵母的有52株；经统计，对三者同时有拮抗作用且作用最强的只有6株，分别为卷曲乳杆菌J3、卷曲乳杆菌J50、卷曲乳杆菌J62、詹氏乳杆菌J87、詹氏乳杆菌J16和格氏乳杆菌J66；不同乳杆菌产生物膜能力数值范围在1.0~5.4，卷曲乳杆菌、詹氏乳杆菌、干酪乳杆菌的生物被膜形成能力显著高于其他三种菌（P<0.05）。在全部179株菌中，卷曲乳杆菌J3和詹氏乳杆菌J87既具有强的产酸能力和产过氧化氢能力，又有较强抑菌活性，同时产生物膜能力也最强。结论 卷曲乳杆菌J3和詹氏乳杆菌J87具有优良的生物学特性，有望成为用于治疗妇科疾病微生态制剂的备选菌株。

Isolation and evaluation of lactic acid bacteria with probiotic characteristics in vagina

Objective To isolate Lactobacillus from healthy female vagina and identify its probiotic properties in order to provide new type of strain for the development of compound probiotics for the treatment of gynecological diseases. Methods Lactobacillus strains were screened from healthy female vaginal secretion and identified by using 16S rDNA sequence analysis. The acid production, H2O2 production, antibacterial activity and biofilm formation were detected. Results From the vaginas of 50 healthy women 179 strains of lactobacillus were isolated, including 101 strains of Lactobacillus crispatus, 42 strains of Lactobacillus jensenii, 26 strains of Lactobacillus gasseri, 5 strains of Lactobacillus plantarum, 3 strains of Lactobacillus saliva and 2 strains of Lactobacillus casei. There were 146 strains of Lactobacillus capable of producing acid. 87 strains were weakly H2O2 positive, 37 strains were H2O2 positive, and 9 strains were strongly H2O2 positive. 115 strains were antagonistic to Escherichia coli, 84 strains were antagonistic to Staphylococcus aureus, while 52 strains were antagonistic to Candida albicans. 6 strains were antagonistic to all the above three bacteria. The value of biofilm forming capacity of these strains ranged from 1.0 to 5.4, among which the biofilm forming ability of Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus casei was significantly higher than the others (P<0.05). In all 179 strains, two strains (Lactobacillus crispatus J3 and Lactobacillus jensenii J87) showed not only strong ability to produce acid and hydrogen peroxide, but also strong antibacterial activity and biofilm forming ability. Conclusion Lactobacillus crispatus J3 and Lactobacillus jensenii J87 are expected to be the candidate strains for the treatment of gynecological diseases.