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Lactobacillus iners and the normal vaginal flora
Linköping University, Department of Clinical and Experimental Medicine, Obstetrics and gynecology. Linköping University, Faculty of Health Sciences.
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [fi]

The ecological niche of the vagina contains a large number of different microbes that are constantly interacting with each other and the host. Culture methods have not been sufficient in order to resolve the complexity of the normal vaginal flora. Further, the methods for delineating normal flora from not normal flora are not easily handled and are traditionally not based on culture but on microscopy of elements of the vaginal fluid. In the work presented in this thesis, an international collaboration was established that pin-pointed some of the difficulties in classifying vaginal floras, including staining, sampling, and discordance when lactobacilli are few in number, and that emphasized the importance of the size of the vision field in microscopes. As lactobacilli are prominent members of the normal vaginal flora they need to be carefully classified if further work towards more robust scoring tools is to be achieved.

Phenotypic methods have not been able to separate the closely related Lactobacillus species of the vagina. Progress in molecular biology has provided possibilities to characterize these lactobacilli, which are mainly from the Lactobacillus acidophilus group. In this work a large number of strains collected by true random sampling were subjected to RAPD-PCR, TTGE and multiplex PCR for species identification. The major species found were L. crispatus, L. gasseri and L. jensenii and the recently described L. iners. The presence of L. iners has not been detected in previous studies due to its special nutrient requirements. Development of pyrosequencing technology also made it possible to match signatures of the two variable regions V1 and V3 of the 16S rRNA gene of the vaginal lactobacilli and identify them to the species level in a high throughput manner. The study confirmed that the dominating flora in women with normal vaginal flora comprises the four species mentioned previously. Repetitive sampling during IVF-treatment with highly varying oestrogen levels demonstrates changes that possibly occur during changes in the natural life cycle. Furthermore, L. iners was found to be the first species to be established after spontaneously resolved or treated Bacterial Vaginosis.

These findings can be of help in developing new strategies for regaining and retaining the normal vaginal flora.

Place, publisher, year, edition, pages
Linköping University Electronic Press, 2008.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1049
Keyword [en]
Vagina, microbes, lactobacilli, phenotypic methods, molecular biology, 16S rRNA, pyrosequencing
National Category
Microbiology in the medical area
Identifiers
URN: urn:nbn:se:liu:diva-11334ISBN: 978-91-7393-953-9 (print)OAI: oai:DiVA.org:liu-11334DiVA: diva2:17743
Public defence
2008-04-18, Berzeliussalen, Campus US, Linköpings universitet, Linköping, 13:00 (English)
Opponent
Supervisors
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2015-11-19
List of papers
1. An international study of the inter-observer variation between the interpretations of vaginal smear criteria of Bacterial Vaginosis
Open this publication in new window or tab >>An international study of the inter-observer variation between the interpretations of vaginal smear criteria of Bacterial Vaginosis
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2002 (English)In: APMIS, ISSN 1600-0463, Vol. 110, no 11, 811-818 p.Article in journal (Refereed) Published
Abstract [en]

An international workshop on vaginal smear-based diagnosis of bacterial vaginosis was organized where 13 investigators scoring 258 slides with smears from vaginal fluid. Interobserver reproducibility of interpretations of Nugent scores, Hay/Ison scores and wet smear scores for the diagnosis of bacterial vaginosis was shown to be high. Detailed analysis of individual scoring results however indicated that basic standards of quality control to ensure robust individual readings of slides must be adhered to.

Keyword
Bacterial vaginosis, diagnosis, criteria
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13050 (URN)10.1034/j.1600-0463.2002.1101107.x (DOI)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2013-09-12
2. Vaginal Lactobacillus flora of healthy Swedish women
Open this publication in new window or tab >>Vaginal Lactobacillus flora of healthy Swedish women
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2002 (English)In: Journal of clinical microbiology, ISSN 0095-1137 (print) 1098-660X (online), Vol. 40, no 8, 2746-2749 p.Article in journal (Refereed) Published
Abstract [en]

Species of the Lactobacillus acidophilus complex are generally considered to constitute most of the vaginal Lactobacillus flora, but the flora varies between studies. However, this may be due to difficulties in identifying the closely related species within the L. acidophilus complex by using traditional methods and to variations in the vaginal status of the participants. Two hundred two isolates from the vaginal fluids of 23 Swedish women without bacterial vaginosis, as defined by the criteria of Nugent et al. (R. P. Nugent, M. A. Krohn, and S. L. Hillier, J. Clin. Microbiol. 29:297-301, 1991), were typed by randomly amplified polymorphic DNA (RAPD) analysis and identified to the species level by temporal temperature gradient gel electrophoresis, multiplex PCR, and 16S ribosomal DNA sequencing. The vaginal flora of most participants was dominated by a single RAPD type, but five of them harbored two RAPD types representing two different species or strains. The most frequently occurring species were Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners, and Lactobacillus jensenii. L. iners has not previously been reported as one of the predominant Lactobacillus species in the vagina.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13051 (URN)10.1128/JCM.40.8.2746-2749.2002 (DOI)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2013-09-12
3. Identification of randomly selected colonies of Lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rDNA Variable V1 and V3 Regions
Open this publication in new window or tab >>Identification of randomly selected colonies of Lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rDNA Variable V1 and V3 Regions
2002 (English)In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 110, no 11, 802-810 p.Article in journal (Refereed) Published
Abstract [en]

The present study aimed to characterize lactobacilli in vaginal fluid from 23 adult healthy women by using high-throughput DNA sequencing for identification of a large number of randomly selected colonies appearing on Rogosa and blood agar. The typing method was based on broad-range PCR of 16S rRNA gene variable regions V1 and V3, pyrosequencing, and classification of the fragments by alignment with NCBI-catalogued sequences and type strain sequences. Four major groups of sequences were found among the 402 isolates clearly corresponding to Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii when compared to the sequences obtained for type strains. Our results indicate that pyrosequencing of 16S rRNA gene fragments as used here is a fast and reliable method well suited for identification to the species level, even within the Lactobacillus acidophilus complex.

Keyword
Lactobacillus, vaginal fluid, 16S rRNA genes, pyrosequencing
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13052 (URN)10.1034/j.1600-0463.2002.1101106.x (DOI)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2016-06-30Bibliographically approved
4. Lactobacillus iners: a marker of changes in the vaginal flora?
Open this publication in new window or tab >>Lactobacillus iners: a marker of changes in the vaginal flora?
2007 (English)In: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 45, no 9, 3145- p.Article in journal, Letter (Other academic) Published
Place, publisher, year, edition, pages
American Society for Microbiology, 2007
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13053 (URN)10.1128/JCM.00558-07 (DOI)000249506900072 ()
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2012-11-01
5. The predominant Human vaginal Lactobacillus flora during IVF treatment
Open this publication in new window or tab >>The predominant Human vaginal Lactobacillus flora during IVF treatment
2008 (English)In: Journal of clinical microbiology, ISSN 0095-1137 (print), 1098-660X (online)Article in journal (Refereed) Submitted
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13054 (URN)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2009-08-19

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