Larry J. Forney! University of Idaho!

Size: px

Start display at page:

Download "Larry J. Forney! University of Idaho!"

Maryann Byrd
5 years ago
Views:

1 Larry J. Forney! University of Idaho!

2 Lactobacillus spp. are characteristic of vaginal microbiota in normal healthy reproductive age women. Growth of non-indigenous organisms, including pathogens, is restricted. The mechanisms are unknown: low ph ( 4.5) lactate other organic acids hydrogen peroxide (?) bacteriocins others?

3 Birth! Rapidly colonized!! Puberty! Increased estrogen; colonization and succession!! Reproductive age! Glycogen metabolized to produce lactic acid!! Menopause! Decreased estrogen, less glycogen, less lactic acid

4 u u u u u BV is the most common cause of vaginal symptoms prompting medical evaluation.! BV is characterized by decreased numbers of lactic acid producing bacteria, elevated vaginal ph, and a disturbed microbiota.! Diagnosis based on clinical criteria or Gram stain (Nugent score).! Numerous epidemiological studies have associated BV with major adverse outcomes, including increase risk for acquiring STIs, including HIV, and pre-term labor.! Not all women are at equal risk to experience BV.! Hypothesis: Differential risk can be accounted for by! differences in the composition, structure, function and! dynamics of their vaginal communities.!

5 u What kinds of bacterial communities are found in vaginas of asymptomatic, healthy women?! u Are these communities dynamic, such that the species composition and evenness varies over time?! u What factors determine vaginal community composition?! u Are changes in community composition associated with increased risk to disease?!

6 A cross-sectional study!

u 410 asymptomatic healthy women! u Four ethnic groups equally represented:!

- Vaginal ph measurements (Inverness VpH glove)! - Nugent scores (used for diagnosis of BV)!

u Determine bacterial community composition by 454 pyrosequencing of barcoded V1-V2 region

7 u 410 asymptomatic healthy women! u Four ethnic groups equally represented:! - Caucasian, Black, Hispanic, Asian! u Self-collected mid-vaginal swabs! u Metadata! - Vaginal ph measurements (Inverness VpH glove)! - Nugent scores (used for diagnosis of BV)! - Questionnaire (health and sexual history, hygiene, etc.)! u Determine bacterial community composition by 454 pyrosequencing of barcoded V1-V2 region of 16S rrna genes! - Assign Lactobacillus sequences to species level! Species level classification using speciateit (speciateit.sourceforge.net)

0 20 40 60 80 % abundance 100 Community State Types!

Community State Types L. iners L. crispatus L. gasseri L.

Eggerthella Rhodobaca Anaerotruncus Ureaplasma Mycoplasma Aerococcus Parvimonas

IV These community states form five distinct clusters - multiple kinds of core

8 % abundance 100 Community State Types! Represent 95% of healthy women in these ethnic groups.! " "- 5% are different! Community State Types L. iners L. crispatus L. gasseri L. jensenii Megasphaera Sneathia Streptococcus Dialister Lachnospira Anaerococcus Eggerthella Rhodobaca Anaerotruncus Ureaplasma Mycoplasma Aerococcus Parvimonas Staphylococcus Corynebacterium Veillonella L.vaginalis II III I V Community States! IV These community states form five distinct clusters - multiple kinds of core microbiomes.! All include significant numbers of lactic acid bacteria, though not necessarily species of Lactobacillus. Function may be conserved.!

9 Principle component analysis shows distribution of states in 3-D space! State type V (L. jensenii) Community states dominated by Lactobacillus spp. are found on the four outer vertices (45%)! State Type IV State type II (L. gasseri) Community states with higher species evenness are found at an inner vertex within the tetrahedron (30%)! Intermediate forms and found along the edges of the tetrahedron (25%)! State type I (L. crispatus) State type III (L. iners) Graphics generated with invue (invue.sourceforge.net)

10 !!! Dynamic equilibrium hypothesis - The composition of a community is somewhat invariant over time and exists in a single dynamic equilibrium.! Community space hypothesis - Communities can occupy any position in community space over time and throughout a woman s lifetime.! Community resilience hypothesis - Communities may vary in composition but are resilient and return to a single community state! Alternative equilibrium states hypothesis - Communities composition can change over time, but the number of alternative states is limited! Longitudinal studies of the vaginal microbiota

11 A longitudinal study!

u 39 women were enrolled in (2004) as part of another study done at Johns Hopkins, Baltimore, MD! u Self-collected swabs were obtained twice-weekly for 16 weeks (total of 1,107 samples)!

12 u 39 women were enrolled in (2004) as part of another study done at Johns Hopkins, Baltimore, MD! u Self-collected swabs were obtained twice-weekly for 16 weeks (total of 1,107 samples)! u Vaginal smears and daily diaries mailed to the lab weekly! u Nugent Gram stain score (0-10 scale)! u 56% African-American and 36% white women! u Analyze community composition by 454 pyrosequencing of barcoded V1-V2 region of 16S rrna genes (>3.5 million reads)!

gasseri Ureaplasma Lactobacillales.2 40 20 0 0.12 D JS t 0.08 0.04 0.

13 Subject 7 Lactobacillales.2 Ureaplasma L.gasseri Lactobacillales.3 Staphylococcus L.jensenii L.unclassified L.crispatus Phylotype L.crispatus L.unclassified L.jensenii Staphylococcus Lactobacillales.3 L.gasseri Ureaplasma Lactobacillales D JS t Menses Tampon Douching Nugent [low,intermediate,high] Vaginal Intercourse Anal Sex Oral Sex Digital Penetration Sex Toy Lubricant weeks

Subject 29 Porphyromonas Streptococcus Gardnerella Aerococcus Anaerococcus L.unclassified L.gasseri 100 80 60 Phylotype L.gasseri L.

14 Subject 29 Porphyromonas Streptococcus Gardnerella Aerococcus Anaerococcus L.unclassified L.gasseri Phylotype L.gasseri L.unclassified Anaerococcus Aerococcus Gardnerella Streptococcus Porphyromonas % % weeks Menses Tampon Douching D JS t Nugent [low,intermediate,high] Vaginal Intercourse Anal Sex Oral Sex Digital Penetration Sex Toy Lubricant

15 Subject 3 Aurantimonadaceae.1 Streptococcus Facklamia Campylobacter Subject 17 Parvimonas Eggerthella L.gasseri Anaerococcus Mobiluncus Megasphaera Aerococcus Gardnerella L.crispatus Corynebacterium Phylotype Corynebacterium L.crispatus Anaerococcus L.gasseri Campylobacter Facklamia Streptococcus Aurantimonadaceae Phylotype Gardnerella Aerococcus Megasphaera Mobiluncus Eggerthella Parvimonas % Menses Tampon Douching D JS t Nugent [low,intermediate,high] Vaginal Intercourse Anal Sex Oral Sex Digital Penetration Sex Toy Lubricant Menses Tampon Douching D JS t Nugent [low,intermediate,high] Vaginal Intercourse Anal Sex Oral Sex Digital Penetration Sex Toy Lubricant weeks weeks

16 Subject 21 Subject 11 Escherichia Streptococcus L.gasseri Gemella Eggerthella Megasphaera Shigella Aerococcus L.jensenii Gemella Eggerthella Aerococcus Megasphaera Gardnerella Ruminococcaceae.1 Sneathia Phylotype L.jensenii Aerococcus Shigella Megasphaera Eggerthella Gemella L.gasseri Streptococcus Escherichia Phylotype Sneathia Ruminococcaceae.1 Gardnerella Megasphaera Aerococcus Eggerthella Gemella %% % % %%%% % % % % % weeks Menses Tampon Douching D JS t Nugent [low,intermediate,high] Vaginal Intercourse Anal Sex Oral Sex Digital Penetration Sex Toy Lubricant D JS t % % % weeks Menses Tampon Douching Nugent [low,intermediate,high] Vaginal Intercourse Anal Sex Oral Sex Digital Penetration Sex Toy Lubricant

17 Vaginal community dynamics Subject 404, Age 45, Monogamous: Yes, Contraceptive: tubal ligation, Male sex partner: Yes Lactobacillaceae.1 Porphyromonas L.unclassified Facklamia Dialister Actinomyces Bacteroides Varibaculum Arcanobacterium Fastidiosipila Propionimicrobium Incertae_Sedis_XI.1 Actinobaculum Campylobacter Parvimonas Corynebacterium Streptococcus Veillonella Anaerococcus L.crispatus ergence 0.0e+00 Ta AnalSex OralSex SexToy weeks

18 Stability versus Constancy!

Differences in Vaginal Bacterial Communities of Women in North America: Implications for disease diagnosis and prevention

Differences in Vaginal Bacterial Communities of Women in North America: Implications for disease diagnosis and prevention Larry J. Forney, Pawel Gajer, Christopher J. Williams, Maria G. Schneider, Stacey