Persistence of Lactobacillus fermentum RC-14 and Lactobacillus rhamnosus GR-1 but Not L. rhamnosus GG in the Human Vagina as Demonstrated by Randomly Amplified Polymorphic DNA

Abstract

Lactobacillus rhamnosus GR-1 and L. fermentum RC-14 are well-characterized probiotic strains with efficacy in the prevention and treatment of urogenital infections in women. The aim of the present study was to apply a molecular biology-based methodology for the detection of these strains and L. rhamnosus GG (a commercially available intestinal probiotic) in the human vagina in order to assess probiotic persistence at this site. Ten healthy women inserted vaginally a capsule containing either a combination of strains GR-1 and RC-14 or the GG strain for 3 consecutive nights. Vaginal swabs taken before and at various time points after probiotic insertion were analyzed, and the Lactobacillus flora was assessed by randomly amplified polymorphic DNA (RAPD) analysis. This method generated discrete DNA fingerprints for GR-1, RC-14, and GG and enabled successful detection of these strains in the vagina. Strain GR-1 and/or strain RC-14 was found to persist in the vaginal tract for up to 19 days after vaginal instillation, while L. rhamnosus GG was detectable for up to 5 days postadministration. In conclusion, the fates of probiotic L. rhamnosus and L. fermentum strains were successfully monitored in the human vagina by RAPD analysis. This technique provides molecular biology-based evidence that RC-14 and GR-1, strains selected as urogenital probiotics, persist in the human vagina and may be more suited to vaginal colonization than L. rhamnosus GG. This highlights the importance of proper selection of strains for urogenital probiotic applications.The vaginal ecosystem is a complex environment in which lactobacilli are the most predominant bacterial species in healthy women (30). A recent study showed that this microflora changes constantly, with only 22% of women having a normal Lactobacillus-dominated flora at any given time (27). As depletion of vaginal lactobacilli can have serious consequences, including increasing the risk of preterm labor and susceptibility to infections such as Neisseria gonorrhoeae, Trichomonas vaginalis, human immunodeficiency virus, and urinary tract infections, bacterial vaginosis, and yeast vaginitis (2,28,30), restoration and maintenance of a normal microflora are important.While antibiotics can be effective for eradication of urinary tract infections, they are often unable to cure bacterial vaginosis or prevent complications that arise from it (6). Furthermore, antibiotics have many side effects, multidrug-resistant microorganisms are common, and such chemotherapeutic agents ideally should not be used for prophylaxis or health maintenance.The use of probiotics, defined as "microbial cell preparations or components of microbial cells that have a beneficial effect on health and well being of the host" (25), offers a potential alternative approach to health restoration and maintenance in the vaginal tract (2,16,29). The question becomes which probiotic strains will persist in and colonize the vagina, given that the vaginal microflora fluctuates daily in terms of both the types and the numbers of...

DNA Fingerprinting of Lactobacillus crispatus Strain CTV-05 by Repetitive Element Sequence-Based PCR Analysis in a Pilot Study of Vaginal Colonization

“…According to Zhong et al, ribotyping appears to distinguish isolates only at the species level (46). More recently, Gardiner et al used random amplified polymorphic DNA-PCR to track three probiotic strains (8). However, it was necessary for them to implement pulsedfield gel electrophoresis to distinguish their L. rhamnosus probiotic strain from closely related strains of the species.…”

Potential Uses of Probiotics in Clinical Practice

“…For strain typing, pulsed-field gel electrophoresis is the gold standard, but randomly amplified polymorphic DNA can also be used (43). Determination of the presence of extrachromosomal genetic elements such as plasmids can also contribute to strain typing and characterization (117).…”

Influence of carbohydrates on cell properties of Lactobacillus rhamnosus

“…The use of lactobacilli as probiotics offers a potential alternative approach to health restoration and maintenance in the vaginal tract. Several representatives of this genus, like Lactobacillus rhamnosus GR-1™ and Lactobacillus fermentum RC-14 have been shown efficient in treatment of bacterial vaginosis [7,8]. The ability of lactobacilli to adhere to epithelial cells is considered as an important feature in the process of colonization of the different human ecological niches such as the vagina.…”