Lactobacillus rhamnosus and Lactobacillus casei Affect Various Stages of Gardnerella Species Biofilm Formation.
Lactobacillus rhamnosus and Lactobacillus casei Affect Various Stages of Gardnerella Species Biofilm Formation.
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DOI：
10.3389/fcimb.2021.568178
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发表时间：
2021
影响因子：
5.7
通讯作者：
Yang H
中科院分区：
文献类型：
--
作者：
He Y;Na R;Niu X;Xiao B;Yang H
细菌性阴道炎（BV）及其复发最常与加德纳菌属生物被膜的形成有关。益生菌通常用于治疗BV；然而，在BV治疗中应用乳酸杆菌益生菌的最佳时期仍不明确。本研究旨在探究鼠李糖乳杆菌和干酪乳杆菌对加德纳菌属生物被膜形成不同阶段的影响。通过结晶紫染色试验测定了7株菌（包括1株阴道加德纳菌ATCC 14018和6株临床分离的加德纳菌属菌株）的生物被膜形成能力。此外，采用微量肉汤稀释法评估浮游菌和生物被膜菌对甲硝唑和克林霉素的敏感性。在加德纳菌属生物被膜形成的不同阶段加入鼠李糖乳杆菌Xbb - LR - 1和干酪乳杆菌Xbb - LC - 1，并共培养24小时。通过共聚焦激光扫描显微镜（CLSM）测定每个样本的生物被膜厚度。通过实时聚合酶链反应法获得每个样本中加德纳菌属的绝对数量，并使用pH试纸测定pH值。通过结晶紫染色、CLSM和扫描电子显微镜（SEM）观察加德纳菌属在不同pH值培养基中的生物被膜形成情况。生物被膜增加了加德纳菌属对甲硝唑和克林霉素的耐药性。在加德纳菌属生物被膜形成初期加入鼠李糖乳杆菌的抑制效果最佳，抑制率为38.17%±1.35%。当培养基的pH值<4.5或>6.5时，ATCC 14018几乎无法形成生物被膜；然而，在pH值≥4.5且≤6.5时，它能够形成更强的生物被膜。在光密度为3.29±0.28（595纳米）、pH值为5.5、培养36小时时，生物被膜量达到最大值。生物被膜的形成增加了加德纳菌属对抗生素的耐药性。维持pH值<4.5的酸性阴道环境以及以乳酸杆菌为主的阴道微生物群，可显著在初始阶段阻止加德纳菌属生物被膜的形成，这对生物被膜相关感染的治疗和预防具有重要影响。
Bacterial vaginosis (BV) and its recurrence are most commonly associated with the formation of Gardnerella species biofilm. Probiotics are typically used to treat BV; however, the optimal period of Lactobacillus probiotic application in BV treatment remains uncertain. The present study aimed to explore the effects of Lactobacillus rhamnosus and Lactobacillus casei on various stages of biofilm formation in Gardnerella species. The biofilm-forming ability of seven strains, including one Gardnerella vaginalis ATCC 14018 and six clinically isolated Gardnerella species, was determined via gentian violet staining assay. Moreover, the sensitivity of the planktonic and biofilm forms toward metronidazole and clindamycin was assessed via microdilution broth method. L. rhamnosus Xbb-LR-1 and L. casei Xbb-LC-1 were added during various stages of biofilm formation in Gardnerella species and were cocultured for 24 h. The biofilm thickness of each sample was determined via confocal laser scanning microscopy (CLSM). The absolute quantities of Gardnerella species in each sample was obtained via real time polymerase chain reaction method, and the pH value was obtained using a pH indicator paper. Biofilm formation by Gardnerella species in a medium with distinct pH values was observed via gentian violet staining, CLSM, and scanning electron microscopy (SEM). The biofilm increased the resistance of Gardnerella species toward metronidazole and clindamycin. L. rhamnosus added at the initial biofilm formation stage in Gardnerella species exhibited highest inhibitory effect, with a percentage inhibition of 38.17% ± 1.35%. When the pH value of the culture medium was <4.5 or >6.5, ATCC 14018 could hardly form a biofilm; however, at pH ≥4.5 and ≤6.5, it was able to form a stronger biofilm. The amount of biofilm attained maximum value at optical density of 3.29 ± 0.28 (595 nm), pH 5.5, and at 36 h. Biofilm formation increases the resistance of Gardnerella species toward antibiotics. Maintaining an acidic vaginal environment with pH <4.5 and a vaginal microbiota dominated by Lactobacillus remarkably prevents the formation of Gardnerella species biofilm at the initial stage, which further has a significant impact on the treatment and prevention of biofilm-related infections.