Fertility and the Microbiome

Fertility and the Microbiome: an Observational Study and Randomized Placebo Controlled Double-blind Pilot Trial

With this study, the investigators want to investigate the microbiome and human papilloma virus (HPV) status of couples with subfertility. The investigators want to gain information about association of female and male microbiome and its impact on fertility. HPV prevalence is high, and its impact on fertility has not been studied intensively. The investigators want to find out whether there is an association between HPV status and subfertility, vaginal and seminal microbiome and HPV status and the prevalence among our subfertile couples.

As part of this study, the investigators will perform a randomized placebo controlled double blind pilot study to investigate the association between altered sperm quality (impaired motility and elevated DNA fragmentation index), the seminal microbiome and whether intake of probiotics alters these parameters.

Study Overview

• Status: Recruiting
• Conditions: Subfertility; Asthenozoospermia
• Intervention / Treatment: Dietary supplement: Probiotics (Omnibiotic (R))

Detailed Description

Observational study:

The investigators will include 150 couples within 24 months and will take samples of oral, urinary, stool, vaginal, uterine and seminal microbiome and cervical as well as seminal samples for HPV analysis. Further, the investigators will test for other sexually transmittable diseases, hormone status and metabolic status. Time to pregnancy, pregnancy rate and live birth rate will be evaluated.

Placebo controlled double blind randomized controlled trial:

30 subfertile men (Asthenozoospermia, elevated DNA fragmentation index) will be randomized in to two groups: Placebo or Probiotics for 3 months daily. Before and after these 3 months, sperm count microbiome analysis and HPV analysis will be performed to find possible associations with intake of probiotics.

• Study Type: Observational
• Enrollment (Estimated): 300

Contacts and Locations

• Study Contact: Name: Sarah Feigl, Dr.; Phone Number: 004331638580831; Email: s.feigl@medunigraz.at

Study Locations

• Austria
  • Styria
    • Graz, Styria, Austria, 8036
      • Recruiting
      • Medical University of Graz
      • Contact: Sarah Feigl, Dr.; Phone Number: 004331638580831; Email: s.feigl@medunigraz.at
      • Contact: Martina Kollmann, PD Dr.; Phone Number: 004331638584190; Email: martina.kollmann@medunigraz.at

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 50 years (Adult)
• Accepts Healthy Volunteers: No

• Sampling Method: Non-Probability Sample

Study Population

Couples (Males and females) with subfertility, primary or secundary infertility, or recurrent pregnancy loss.

Description

Inclusion Criteria:

• Couples (Males and females) with subfertility, primary or secondary infertility, or recurrent pregnancy loss.

Exclusion Criteria:

• unable to consent
• diseases or conditions that might impact investigation
• intake of probiotics, antibiotics within last 3 months
• intake of nutritional supplements
• chemo therapy or radiation therapy
• excess alcohol intake
• smoking

Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Subfertile patients; Males and females between 18 and 50 years of age with subfertility presenting at the fertility center at the Medical University of Graz.
Subfertile males with elevated DNA fragmentation index and asthenozoospermia; Males between 18 and 50 years of age with elevated DNA fragmentation index and asthenozoospermia presenting at the fertility center at the Medical University of Graz.	Dietary supplement: Probiotics (Omnibiotic (R)); As intervention in the randomized placebo controlled double blind trial, 30 males will receive either probiotics or placebo once daily for 3 months. Sperm count and microbiome analysis before and after intervention will be performed.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Seminal microbiome	16S-RNA sequencing; shotgun metagenomic sequencing	3 months
Vaginal microbiome	16S-RNA sequencing; shotgun metagenomic sequencing	1 month
Uterine microbiome	16S-RNA sequencing; shotgun metagenomic sequencing	1 month
DNA-Fragmentation index	Rate of sperm with elevated DNA Fragmentation index	3 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Hormone status of males and females - LH (Luteinizing hormone)	LH status	3 months
Hormone status of males and females - FSH (follicle stimulating hormone)	FSH status	3 months
Hormone status of males and females - Testosterone status	Testosterone status	3 months
Hormone status of males and females - free Testosterone	free Testosterone status	3 months
Hormone status of males and females - Androstenedione	Androstenedione status	3 months
Hormone status of males and females - AMH (Anti-Müllerian hormone)	AMH status	3 months
Hormone status of males and females - DHEAS (dehydroepiandrosterone sulphate)	DHEAS status	3 months
Hormone status of males and females - 25(OH)Vitamin D	25(OH)Vitamin D status	3 months
Hormone status of males and females - TSH (thyroid-stimulating hormone)	TSH status	3 months
Hormone status of males and females - fT3 (free Triiodothyronine)	fT3 Status	3 months
Hormone status of males and females - freeThyroxine (fT4)	fT4 status	3 months
HPV Status	HPV PCR analysis	3 months
stool microbiome	16S-RNA sequencing; shotgun metagenomic sequencing	3 months
oral microbiome	16S-RNA sequencing; shotgun metagenomic sequencing	3 months
Cortisol in Saliva	cortisol analysis in saliva of females and males	3 months
Amylase in Saliva	amylase analysis in saliva of females and males	3 months
Metabolic parameters - Omega 3	Omega 3	3 months
Metabolic parameters - Omega 6	Omega 6	3 months
Metabolic parameters - low density lipoprotein	low density lipoprotein	3 months
Metabolic parameters - high density lipoprotein	high density lipoprotein	3 months
Metabolic parameters - triglycerides	triglycerides	3 months
Metabolic parameters - LipoproteinA	LipoproteinA	3 months
Time to pregnancy	Time until clinical pregnancy is achieved	24 months
pregnancy rate	rate of clinical pregnancies	24 months
live birth rate	follow up of pregnancies and their outcome	24 months
Presence of sexual transmittable diseases	HBV, HCV, HIV, Syphilis, Chlamydia, Gardnerella, Trichomonas	3 months
Sperm count	Motility, concentration, morphology	3 months

Collaborators and Investigators

• Sponsor: Medical University of Graz
• Investigators: Principal Investigator: Martina Kollmann, PD Dr., Medical University of Graz

Publications and helpful links

General Publications

• Baud D, Pattaroni C, Vulliemoz N, Castella V, Marsland BJ, Stojanov M. Sperm Microbiota and Its Impact on Semen Parameters. Front Microbiol. 2019 Feb 12;10:234. doi: 10.3389/fmicb.2019.00234. eCollection 2019.
• Jersoviene V, Gudleviciene Z, Rimiene J, Butkauskas D. Human Papillomavirus and Infertility. Medicina (Kaunas). 2019 Jul 15;55(7):377. doi: 10.3390/medicina55070377.
• Brusselaers N, Shrestha S, van de Wijgert J, Verstraelen H. Vaginal dysbiosis and the risk of human papillomavirus and cervical cancer: systematic review and meta-analysis. Am J Obstet Gynecol. 2019 Jul;221(1):9-18.e8. doi: 10.1016/j.ajog.2018.12.011. Epub 2018 Dec 12.
• Koedooder R, Singer M, Schoenmakers S, Savelkoul PHM, Morre SA, de Jonge JD, Poort L, Cuypers WJSS, Beckers NGM, Broekmans FJM, Cohlen BJ, den Hartog JE, Fleischer K, Lambalk CB, Smeenk JMJS, Budding AE, Laven JSE. The vaginal microbiome as a predictor for outcome of in vitro fertilization with or without intracytoplasmic sperm injection: a prospective study. Hum Reprod. 2019 Jun 4;34(6):1042-1054. doi: 10.1093/humrep/dez065. Erratum In: Hum Reprod. 2019 Oct 2;34(10):2091-2092.
• Depuydt CE, Donders GGG, Verstraete L, Vanden Broeck D, Beert JFA, Salembier G, Bosmans E, Ombelet W. Infectious human papillomavirus virions in semen reduce clinical pregnancy rates in women undergoing intrauterine insemination. Fertil Steril. 2019 Jun;111(6):1135-1144. doi: 10.1016/j.fertnstert.2019.02.002. Epub 2019 Apr 17.
• Koedooder R, Singer M, Schoenmakers S, Savelkoul PHM, Morre SA, de Jonge JD, Poort L, Cuypers WSS, Budding AE, Laven JSE; ReceptIVFity study group. The ReceptIVFity cohort study protocol to validate the urogenital microbiome as predictor for IVF or IVF/ICSI outcome. Reprod Health. 2018 Dec 7;15(1):202. doi: 10.1186/s12978-018-0653-x.
• Depuydt CE, Donders G, Verstraete L, Vanden Broeck D, Beert J, Salembier G, Bosmans E, DhontT N, Van Der Auwera I, Vandenborne K, Ombelet W. Time has come to include Human Papillomavirus (HPV) testing in sperm donor banks. Facts Views Vis Obgyn. 2018 Dec;10(4):201-205.
• Xiong YQ, Chen YX, Cheng MJ, He WQ, Chen Q. The risk of human papillomavirus infection for male fertility abnormality: a meta-analysis. Asian J Androl. 2018 Sep-Oct;20(5):493-497. doi: 10.4103/aja.aja_77_17.
• Moreno I, Simon C. Relevance of assessing the uterine microbiota in infertility. Fertil Steril. 2018 Aug;110(3):337-343. doi: 10.1016/j.fertnstert.2018.04.041.
• Simon C. Introduction: Do microbes in the female reproductive function matter? Fertil Steril. 2018 Aug;110(3):325-326. doi: 10.1016/j.fertnstert.2018.06.041. Epub 2018 Jul 20.

Study record dates

Study Major Dates

• Study Start (Actual): August 10, 2022
• Primary Completion (Estimated): April 11, 2024
• Study Completion (Estimated): December 31, 2024

Study Registration Dates

• First Submitted: April 7, 2022
• First Submitted That Met QC Criteria: April 7, 2022
• First Posted (Actual): April 14, 2022

Study Record Updates

• Last Update Posted (Actual): October 23, 2023
• Last Update Submitted That Met QC Criteria: October 18, 2023
• Last Verified: October 1, 2023

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Infertility, Male; Urogenital Diseases; Male Urogenital Diseases; Genital Diseases, Male; Genital Diseases; Infertility; Asthenozoospermia
• Other Study ID Numbers: Fertilitom

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No