Poster presented by Roxana Hickey (PhD candidate, University of Idaho) at #ISME15 in Seoul, South Korea. Session PS05, board 098A. On display Monday 8/25—Tuesday 8/26.

1. Vaginal microbiota of adolescent girls resemble those of
reproductive-age women prior to the onset of menarche
Roxana Hickey1,2, Xia Zhou1, Matthew Settles2, Julie Erb3, Kristin Malone3,
Melanie Hansmann3, Barbara van der Pol4, J. Dennis Fortenberry5 & Larry Forney1,2
1Department of Biological Sciences & 2Institute for Bioinformatics and Evolutionary Studies, University of Idaho, Moscow, ID, USA; 3FemCare Division, Procter &
Gamble, Cincinnati, OH, USA; 4Indiana University School of Public Health, Bloomington, IN, USA; 5Indiana University School of Medicine, Indianapolis, IN, USA
introduction
The vaginal microbiome over a lifespan
๏ During reproductive-age years, vaginal
microbiota differ among women1 and are
variable over short time scales.2
๏ Lactic acid bacteria (esp. Lactobacillus) and
maintenance of low vaginal pH (~4.5) are
considered hallmark conditions of health.
๏ Little is known about the timing and sequence
of changes that occur during puberty, but
microbiota of postmenarcheal teens (13-18 y/o)
appear to resemble those of adults.3,4
๏ Studying transitions during puberty is an
important first step toward understanding
community assembly and succession in healthy
vaginal microbiota.
methods
Study design
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Longitudinal patterns differed considerably among individuals, but all had
abundant lactic acid bacteria prior to or at menarche. Vaginal microbiota of
girls and their mothers were only occasionally similar.
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Subject 101 Subject 102 Subject 103 Subject 104
Subject 105 Subject 106 Subject 107 Subject 108
Subject 109 Subject 110 Subject 111 Subject 112
Subject 113 Subject 114 Subject 115 Subject 116
Subject 118 Subject 120 Subject 121 Subject 123
Subject 124 Subject 125 Subject 126 Subject 127
Subject 128 Subject 129 Subject 132 Subject 133
Subject 134 Subject 135 Subject 136
Inconvenient truth: girls have Gardnerella, too
๏ Commonly associated with bacterial vaginosis in adults,
Gardnerella vaginalis was detected in 11/31 girls.
๏ Current efforts underway to explore the genetic diversity
of this species across different age groups: adolescent
girls, reproductive-age and postmenopausal women.
Vaginal swabs collected every
3 months for up to 3 years
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acknowledgments
We are grateful for support and advice from Dorah Mtui, Daniel Beck, Matt Pennell
and Sam Hunter (U Idaho); Alexis Roth, Cynthia Robbins, and Marcia Shew
(Indiana U SoM); and Lisa Bowman (P&G). Financial support was provided by
Procter & Gamble, NIH P30 RR033376, and the UI BCB Graduate Program.
Girl Vagina
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references
1Ravel et al. (2011) PNAS 108:S1. 2Gajer et al. (2012) Sci Transl Med 4.
3Yamamoto et al. (2009) J Pediatr Adolesc Gynecol 22. 4Thoma et al. (2010) J
Pediatr Adolesc Gynecol 1. 5Schloss et al. (2009) Appl Environ Microbiol 75.
6Cole et al. (2005) Nuc Acids Res 33. 7http://patricbrc.org/.
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more information
Forney lab: http://people.ibest.uidaho.edu/~lforney/
roxana.hickey@gmail.com lforney@uidaho.edu
results
conclusions & next steps
Unexpectedly, vaginal microbiota of girls
were often characterized by a dominance
of lactic acid bacteria prior to menarche.
๏ Bacterial composition:
Roche 454 sequencing
of 16S rRNA genes
(V1-V3)
‣ mothur5
‣ RDP Naïve Bayesian
Classifier6
‣ PATRIC7 for speciation
of Lactobacillus spp.
๏ Metadata: Tanner stage,
vaginal pH, menarcheal
status
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Figure 1. Tanner stages of
breast and pubic development.
Lactic acid bacteria became
abundant early in puberty, while
vaginal pH declined more slowly.
Figure 2. Bacterial community composition of the vaginal microbiota of
girls and mothers sampled longitudinally. 198 samples from 31 girls and
47 samples from 24 mothers are represented. The dendrogram represents
the average linkage clustering based on the Bray-Curtis dissimilarity
computed from Hellinger standardized taxon abundance data. The
heatmap depicts proportions of the 25 overall most abundant taxa.
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2 3 4 5
Tanner breast
Logit−transformed LAB proportion
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−5
pre post
Menarche status
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2 3 4 5
Tanner breast
Vaginal pH
4
pre post
Menarche status
Figure 3. Trends in relative abundance of lactic
acid bacteria and vaginal pH in relation to
pubertal development and menarche status. Box
plots show (a) logit-transformed proportions of
lactic acid bacteria (LAB) and (b) vaginal pH.
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Proportion
Subject 107
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Subject 109
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Proportion
Subject 102
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Subject 103
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Visit
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Visit
Taxon
Lactobacillus_crispatus
Lactobacillus_iners
Lactobacillus_gasseri
Gardnerella_vaginalis
Lactobacillus_jensenii
Lactobacillus_Other
Streptococcus_anginosis
Bifidobacterium
Gardnerella_Other
Prevotella
Anaerococcus
Other_Bacteria
Atopobium
Finegoldia
Peptoniphilus
Ureaplasma
Bacteroides
Dialister
Faecalibacterium
Corynebacterium
Lachnospiracea_incertae_sedis
Alloscardovia
Moryella
Actinomyces
Clostridiales_Incertae_Sedis_XI
Porphyromonas
Blautia
Clostridiales
Anaerosphaera
Jonquetella
Mobiluncus
Actinomycetales
Staphylococcus
Segniliparus
Clostridium_XI
Lachnospiraceae
Coriobacteriaceae
Acidovorax
Olsenella
Peptococcus
Tanner breast ●1●2●3●4●5
Menarche status ●Pre●Post
Figure 5. Transitions to Lactobacillus-dominant microbiota in
four girls. Panels show the vaginal bacterial community profiles
and associated pubertal development of four participants
sampled longitudinally. Bar plots represent the proportions of
bacterial taxa in the community. Menarcheal status and Tanner
breast stage are indicated below each bar plot.
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Mother Vagina
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Mother Vagina
Mother Vagina
Mother Vagina
Mother Vagina
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Visit No.
Cluster
Group
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Bifido
GV
LC
LG
LI
LJ
Other
Sample
Type
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Post
Mother
Figure 4. Hierarchical cluster assignment over time within
individual participants. Each plot shows the hierarchical cluster
assignment (as in Figure 2) of vaginal microbiota samples from a
girl (circles) and her mother (triangles), when applicable. The x-axis
indicates the clinical visit at which each sample was collected
(occurred approximately every three months).
31 premenarcheal
10-12 y/o girls
‣ 21 Black
‣ 8 Caucasian
‣ 2 Native American
21 girls became
postmenarcheal
during study
24 mothers
sampled annually
Common trends, divergent details
๏ Transitions to ‘adult-like' microbiota appear to occur in the
early to intermediate stages of puberty before menarche begins.
๏ As in adults, vaginal microbiota of girls vary both among and
within individuals. This may be essential to understanding
differences in ecological function and stability of microbiota.
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Visit No.
Proportion of Gardnerella
Menarche
Status
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Post
Figure 6. Proportion of Gardnerella over
time in the vaginal microbiota of girls.
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