Material & Methods

My study was divided into three main parts related to the different biomarkers I used: 1) sperm motility 2) oxidative stress status of seminal plasma 3) metabolite composition of seminal plasma.

Sample size: A total number of 60 ejaculates were collected from 20 young bulls (three ejaculates per individual).

  1. Sperm motility study: Eight motility parameters were studied by Computer-assisted sperm motility analysis (CASA), which uses short video recordings of live samples. Based on these parameters two groups were formed: Low and High quality ejaculates.  
Picture of CASA machine taken from:

2. Oxidative stress status of seminal plasma composition: Concentrations of advanced oxidative protein products (AOPPs), protein carbonylation (carbonyls) were measured, using spectrophotometry. These parameters are indicators of ROS generation.

Concentration of thiols a type of antioxidant was also measured using spectrophotometry.

3. Metabolite composition of seminal plasma:

Metabolites were studied using  proton nuclear magnetic resonance (1H NMR). NMR determines the atomic composition of metabolites and their concentrations in a given sample.

Picture of NMR machine taken from: