Fertility and the Microbiome

Study Overview
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Brief Summary
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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.

Detailed Description
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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.

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

Conditions
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Subfertility, Asthenozoospermia

Intervention / Treatment
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* Dietary Supplement: Probiotics (Omnibiotic (R))


Participation Criteria
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Eligibility Criteria
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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

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 50 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Subfertile patients<br>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<br>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))<br>* 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.<br>|

What is the study measuring?
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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 |