Probiotics on Sperm Quality in Male Infertility Patients

Content: Male infertility accounts for approximately 40% of infertility cases and is often characterized by abnormalities in sperm count, motility, and morphology. Factors such as oxidative stress, environmental hormones, and inflammation can lead to sperm DNA fragmentation and mitochondrial dysfunction, ultimately affecting fertilization potential. While probiotics are known to improve gut microbiome balance and regulate immunity, limited studies suggest they may also improve sperm motility and reduce DNA fragmentation in men with asthenozoospermia.

This pilot study employs a randomized, placebo-controlled design to evaluate the efficacy of probiotic supplementation in improving sperm quality. A total of 60 male participants with unexplained infertility (specifically oligozoospermia, asthenozoospermia, or oligoasthenoteratozoospermia) will be recruited. Participants will be divided into two groups: an experimental group receiving probiotics and a control group receiving a placebo.

The study duration for each participant is approximately 100 days, with semen samples collected at three distinct time points:

• Day 0 (Start Test): Baseline sample collection.
• Day 60 (Mid Test): Interim sample collection.
• Day 100 (Final Test): Post-intervention sample collection.

Semen analysis will be conducted using the following methodologies:

• Sperm Concentration and Motility: Analyzed using a Computer-Aided Sperm Assay (CASA) to measure parameters including total motility, progressive motility, VAP, VCL, VSL, Straightness, and Linearity.
• Sperm DNA Integrity: Assessed via the Sperm Chromatin Structure Assay (SCSA) using flow cytometry. Sperm are treated with acridine orange; red fluorescence indicates single-stranded DNA (fragmentation), while green fluorescence indicates double-stranded DNA. Results are expressed as the DNA Fragmentation Index (DFI).

Sperm Function (Acrosome Reaction): Evaluated using Peanut Agglutinin (PNA) staining. The assay determines the percentage of sperm capable of undergoing the acrosome reaction, a necessary step for fertilization.

Inclusion criteria

• Gender: Male.
• Age: 20 to 45 years old.
• Diagnosis: Diagnosed with unexplained oligozoospermia, asthenozoospermia, or oligoasthenoteratozoospermia.
• Semen Parameters:

• Sperm concentration < 5×106/mL (Note: Source indicates "5x10/mL", contextually adjusted to standard unit for oligozoospermia).
• Sperm motility < 40%.
• Sperm morphology (Kruger strict criteria) < 4%. Exclusion Criteria

• History of hormonal disorders or epididymo-orchitis.
• Substance abuse, including drugs or excessive alcohol consumption.
• Diabetes mellitus.
• Kidney disease (defined as a doubling of creatinine levels or more).
• Chronic liver disease.
• Varicocele.
• Current use of medications that interfere with hormones.
• Occupational or environmental exposure to pesticides, heavy metals, or solvents.
• Intake of antioxidant supplements within the past three months.
• Body Mass Index (BMI) of 30 kg/m² or higher.

Inclusion criteria

• Males aged 20 to 45 years
• Diagnosis of unexplained oligozoospermia, asthenozoospermia, or oligoasthenoteratozoospermia
• Sperm concentration < 5 x 10^6/mL
• Sperm total motility < 40%
• Sperm morphology (Kruger strict criteria) < 4%

Exclusion criteria

• History of hormonal disorders or epididymo-orchitis
• Substance abuse, including drugs or excessive alcohol consumption
• Diabetes mellitus
• Kidney disease (defined as a doubling of creatinine levels or more)
• Chronic liver disease
• Varicocele
• Current use of medications that interfere with hormones
• Occupational or environmental exposure to pesticides, heavy metals, or solvents
• Intake of antioxidant supplements within the past three months
• Body Mass Index (BMI) of 30 kg/m^2 or higher