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Community dynamics of the
adolescent vaginal
microbiome during puberty
Roxana Hickey
University of Idaho
META Symposium on Host-Microbe Systems Biology
9 August 2014
@roxana_hickey
How does the vaginal microbiome
change over a lifespan?
Birth
Vagina sterile in utero (?), then transiently colonized by
mother’s vagina or skin microbiota upon delivery
Childhood
Vagina sparsely populated by assortment of
anaerobic bacteria, few to no lactobacilli
Adolescence
Estrogen levels rise, lactobacilli become more prevalent
while anaerobic populations diminish
Reproductive age
Estrogen cycling stimulates thickening of vaginal
epithelium, abundant lactobacilli help maintain low pH
Menopause
Estrogen levels decline, vaginal epithelium thins,
lactobacilli diminish, anaerobes prevalent
Vaginal microbiota differ among individuals and
are often variable over short time scales
Ravel et al. (2011) PNAS
Gajer et al. (2012) Sci Transl Med
??? ?
Vaginal microbiota of postmenarcheal adolescents (13-18y)
closely resemble those of reproductive age women
Yamamoto et al. (2009) J Pediatr Adolesc Gynecol
Thoma et al. (2011) J Pediatr Adolesc Gynecol
What is the timing and sequence of events that occur as
vaginal microbiota transition to an ‘adult-like’ state?
Study design
microbiota composition
๏ 16S rRNA V1-V3 pyrosequencing
๏ mothur + RDP Naïve Bayesian
Classifier
24 mothers
sampled annually
31 girls enrolled
@ age 10-12 &
premenarcheal
sampled every 3 months for up to 3 years
21 girls became
postmenarcheal
during study
metadata
๏ pubertal development (Tanner stage)
๏ menarcheal status
๏ 21 Black
๏ 8 Caucasian
๏ 2 Native American
Most adolescent microbiota
were already ‘adult-like’
prior to menarche
Lactobacillus crispatus L. iners
L.gasseri
L.jensenii
Gardnerellavaginalis
‘Other’
Lactic acid bacteria became
prevalent early in puberty
●
●
●●
●
●
●
●
●
●
●
−5
0
5
2 3 4 5
Tanner breast
Logit−transformedLABproportion
a
●
●●
−5
0
5
pre post
Menarche status
b
1
2
3
4
5
Tanner stages
Wikimedia Commons
50%
~100%
~0%
Many paths to the
same (?) end
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 107
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 109
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0.00
0.25
0.50
0.75
1.00
Proportion
Subject 102
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 103
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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
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 107
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 109
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● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11
Visit
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 102
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 103
● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11 12
Visit
● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11 12
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
An inconvenient truth: girls
have Gardnerella, too
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 102
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 120
● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11 12
Visit
● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●
1 2 3 4 5 6
Visit
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 121
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 135
● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●
1 2 3 4 5 6 7 8
Visit
● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●
1 2 3 4 5
Visit
Taxon
Gardnerella_vaginalis
Lactobacillus_iners
Lactobacillus_crispatus
Gardnerella_Other
Lactobacillus_gasseri
Prevotella
Atopobium
Anaerococcus
Lactobacillus_Other
Finegoldia
Sneathia
Peptoniphilus
Lactobacillus_jensenii
Dialister
Other_Bacteria
Ureaplasma
Mobiluncus
Lachnospiraceae
Megasphaera
Coriobacteriaceae
Porphyromonadaceae
Veillonella
Alloscardovia
Clostridiales
Anaerosphaera
Ruminococcaceae
Porphyromonas
Clostridiales_Incertae_Sedis_XI
Jonquetella
Veillonellaceae
Gemella
Corynebacterium
Staphylococcus
Lactobacillus_coleohominis
Aerococcus
Segniliparus
Actinomycetales
Moryella
Escherichia_Shigella
Parvimonas
Tanner breast ● ● ● ● ●1 2 3 4 5
Menarche status ● ●Pre Post
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 102
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 120
● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11 12
Visit
● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●
1 2 3 4 5 6
Visit
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 121
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 135
● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●
1 2 3 4 5 6 7 8
Visit
● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●
1 2 3 4 5
Visit
Taxon
Gardnerella_vaginalis
Lactobacillus_iners
Lactobacillus_crispatus
Gardnerella_Other
Lactobacillus_gasseri
Prevotella
Atopobium
Anaerococcus
Lactobacillus_Other
Finegoldia
Sneathia
Peptoniphilus
Lactobacillus_jensenii
Dialister
Other_Bacteria
Ureaplasma
Mobiluncus
Lachnospiraceae
Megasphaera
Coriobacteriaceae
Porphyromonadaceae
Veillonella
Alloscardovia
Clostridiales
Anaerosphaera
Ruminococcaceae
Porphyromonas
Clostridiales_Incertae_Sedis_XI
Jonquetella
Veillonellaceae
Gemella
Corynebacterium
Staphylococcus
Lactobacillus_coleohominis
Aerococcus
Segniliparus
Actinomycetales
Moryella
Escherichia_Shigella
Parvimonas
Tanner breast ● ● ● ● ●1 2 3 4 5
Menarche status ● ●Pre Post
Same,
but different
Same,
●
●
●●
●
●
●
●
●
●
●
−5
0
5
2 3 4 5
Tanner breast
Logit−transformedLABproportion
a
●
●●
−5
0
5
pre post
Menarche status
●
4
5
6
7
2 3 4 5
Tanner breast
VaginalpH
b
4
5
6
7
pre post
Menarche status
!
but different
!
but different
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 107
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 109
● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11
Visit
● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11
Visit
0.50
0.75
1.00
oportion
Subject 102
0.50
0.75
1.00
oportion
Subject 103
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 107
0.00
0.25
0.50
0.75
1.00
Proportion
Subject 109
● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●
1 2 3 4 5 6 7 8 9 10 11
Visit
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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
Same,
Acknowledgements
Procter & Gamble
FEMCARE DIVISION
Melanie Hansmann
Kristin Malone
Julie Erb
Lisa Bowman
Funding
Procter & Gamble
NIH P30 RR033376
UI BCB Graduate Fellowship
Indiana University
SCHOOL OF MEDICINE
J. Dennis Fortenberry
Alexis Roth
Cynthia Robbins
Marcia Shew
SCHOOL OF PUBLIC HEALTH
Barbara van der Pol
University of Idaho
Larry Forney
Xia Zhou
Matt Settles
Dorah Mtui
Daniel Beck
Matt Pennell
Sam Hunter
Forney & Top Labs
IBEST
@roxana_hickey

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Community dynamics of the adolescent vaginal microbiome during puberty (UOregon META Symposium 2014)

  • 1. Community dynamics of the adolescent vaginal microbiome during puberty Roxana Hickey University of Idaho META Symposium on Host-Microbe Systems Biology 9 August 2014 @roxana_hickey
  • 2. How does the vaginal microbiome change over a lifespan?
  • 3. Birth Vagina sterile in utero (?), then transiently colonized by mother’s vagina or skin microbiota upon delivery
  • 4. Childhood Vagina sparsely populated by assortment of anaerobic bacteria, few to no lactobacilli
  • 5. Adolescence Estrogen levels rise, lactobacilli become more prevalent while anaerobic populations diminish
  • 6. Reproductive age Estrogen cycling stimulates thickening of vaginal epithelium, abundant lactobacilli help maintain low pH
  • 7. Menopause Estrogen levels decline, vaginal epithelium thins, lactobacilli diminish, anaerobes prevalent
  • 8.
  • 9. Vaginal microbiota differ among individuals and are often variable over short time scales Ravel et al. (2011) PNAS Gajer et al. (2012) Sci Transl Med
  • 10. ??? ?
  • 11. Vaginal microbiota of postmenarcheal adolescents (13-18y) closely resemble those of reproductive age women Yamamoto et al. (2009) J Pediatr Adolesc Gynecol Thoma et al. (2011) J Pediatr Adolesc Gynecol
  • 12. What is the timing and sequence of events that occur as vaginal microbiota transition to an ‘adult-like’ state?
  • 13. Study design microbiota composition ๏ 16S rRNA V1-V3 pyrosequencing ๏ mothur + RDP Naïve Bayesian Classifier 24 mothers sampled annually 31 girls enrolled @ age 10-12 & premenarcheal sampled every 3 months for up to 3 years 21 girls became postmenarcheal during study metadata ๏ pubertal development (Tanner stage) ๏ menarcheal status ๏ 21 Black ๏ 8 Caucasian ๏ 2 Native American
  • 14. Most adolescent microbiota were already ‘adult-like’ prior to menarche
  • 15.
  • 16. Lactobacillus crispatus L. iners L.gasseri L.jensenii Gardnerellavaginalis ‘Other’
  • 17. Lactic acid bacteria became prevalent early in puberty
  • 18. ● ● ●● ● ● ● ● ● ● ● −5 0 5 2 3 4 5 Tanner breast Logit−transformedLABproportion a ● ●● −5 0 5 pre post Menarche status b 1 2 3 4 5 Tanner stages Wikimedia Commons 50% ~100% ~0%
  • 19. Many paths to the same (?) end
  • 20. 0.00 0.25 0.50 0.75 1.00 Proportion Subject 107 0.00 0.25 0.50 0.75 1.00 Proportion Subject 109 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit 0.00 0.25 0.50 0.75 1.00 Proportion Subject 102 0.00 0.25 0.50 0.75 1.00 Proportion Subject 103 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 Visit ● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 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
  • 21. 0.00 0.25 0.50 0.75 1.00 Proportion Subject 107 0.00 0.25 0.50 0.75 1.00 Proportion Subject 109 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit 0.00 0.25 0.50 0.75 1.00 Proportion Subject 102 0.00 0.25 0.50 0.75 1.00 Proportion Subject 103 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 Visit ● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 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
  • 22. An inconvenient truth: girls have Gardnerella, too
  • 23. 0.00 0.25 0.50 0.75 1.00 Proportion Subject 102 0.00 0.25 0.50 0.75 1.00 Proportion Subject 120 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 Visit ● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ● 1 2 3 4 5 6 Visit 0.00 0.25 0.50 0.75 1.00 Proportion Subject 121 0.00 0.25 0.50 0.75 1.00 Proportion Subject 135 ● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ● 1 2 3 4 5 6 7 8 Visit ● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ● 1 2 3 4 5 Visit Taxon Gardnerella_vaginalis Lactobacillus_iners Lactobacillus_crispatus Gardnerella_Other Lactobacillus_gasseri Prevotella Atopobium Anaerococcus Lactobacillus_Other Finegoldia Sneathia Peptoniphilus Lactobacillus_jensenii Dialister Other_Bacteria Ureaplasma Mobiluncus Lachnospiraceae Megasphaera Coriobacteriaceae Porphyromonadaceae Veillonella Alloscardovia Clostridiales Anaerosphaera Ruminococcaceae Porphyromonas Clostridiales_Incertae_Sedis_XI Jonquetella Veillonellaceae Gemella Corynebacterium Staphylococcus Lactobacillus_coleohominis Aerococcus Segniliparus Actinomycetales Moryella Escherichia_Shigella Parvimonas Tanner breast ● ● ● ● ●1 2 3 4 5 Menarche status ● ●Pre Post
  • 24. 0.00 0.25 0.50 0.75 1.00 Proportion Subject 102 0.00 0.25 0.50 0.75 1.00 Proportion Subject 120 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 Visit ● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ●● ● ● ● ● 1 2 3 4 5 6 Visit 0.00 0.25 0.50 0.75 1.00 Proportion Subject 121 0.00 0.25 0.50 0.75 1.00 Proportion Subject 135 ● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ●● ● ● ● ● ● 1 2 3 4 5 6 7 8 Visit ● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ●● ● ● ● 1 2 3 4 5 Visit Taxon Gardnerella_vaginalis Lactobacillus_iners Lactobacillus_crispatus Gardnerella_Other Lactobacillus_gasseri Prevotella Atopobium Anaerococcus Lactobacillus_Other Finegoldia Sneathia Peptoniphilus Lactobacillus_jensenii Dialister Other_Bacteria Ureaplasma Mobiluncus Lachnospiraceae Megasphaera Coriobacteriaceae Porphyromonadaceae Veillonella Alloscardovia Clostridiales Anaerosphaera Ruminococcaceae Porphyromonas Clostridiales_Incertae_Sedis_XI Jonquetella Veillonellaceae Gemella Corynebacterium Staphylococcus Lactobacillus_coleohominis Aerococcus Segniliparus Actinomycetales Moryella Escherichia_Shigella Parvimonas Tanner breast ● ● ● ● ●1 2 3 4 5 Menarche status ● ●Pre Post
  • 26. Same, ● ● ●● ● ● ● ● ● ● ● −5 0 5 2 3 4 5 Tanner breast Logit−transformedLABproportion a ● ●● −5 0 5 pre post Menarche status ● 4 5 6 7 2 3 4 5 Tanner breast VaginalpH b 4 5 6 7 pre post Menarche status ! but different
  • 27. ! but different 0.00 0.25 0.50 0.75 1.00 Proportion Subject 107 0.00 0.25 0.50 0.75 1.00 Proportion Subject 109 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit 0.50 0.75 1.00 oportion Subject 102 0.50 0.75 1.00 oportion Subject 103 0.00 0.25 0.50 0.75 1.00 Proportion Subject 107 0.00 0.25 0.50 0.75 1.00 Proportion Subject 109 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 Visit 0.00 0.25 0.50 0.75 1.00 Proportion Subject 102 0.00 0.25 0.50 0.75 1.00 Proportion Subject 103 ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 Visit ● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ●● ● ● ● ● ● ● ● ● ● ● ● 1 2 3 4 5 6 7 8 9 10 11 12 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 Same,
  • 28. Acknowledgements Procter & Gamble FEMCARE DIVISION Melanie Hansmann Kristin Malone Julie Erb Lisa Bowman Funding Procter & Gamble NIH P30 RR033376 UI BCB Graduate Fellowship Indiana University SCHOOL OF MEDICINE J. Dennis Fortenberry Alexis Roth Cynthia Robbins Marcia Shew SCHOOL OF PUBLIC HEALTH Barbara van der Pol University of Idaho Larry Forney Xia Zhou Matt Settles Dorah Mtui Daniel Beck Matt Pennell Sam Hunter Forney & Top Labs IBEST