# Integrating Near-Infrared Spectroscopy and Proteomics for Semen Quality Biosensing

## TL;DR

This study combines near-infrared spectroscopy and proteomics to non-invasively assess boar semen quality using extracellular vesicles in seminal plasma.

## Contribution

The integration of NIRS and SP-EV proteomics offers a novel, non-invasive method for rapid semen quality evaluation in swine breeding.

## Key findings

- NIRS achieved 92.2% accuracy in classifying semen quality based on water structure variations.
- Proteomic analysis identified 218 and 238 proteins in passed and failed samples, respectively, with key proteins like Nexin-1 and LGALS3BP showing differential expression.
- Functional analysis linked passed samples to pathways involving fertilization and glutathione metabolism.

## Abstract

Artificial insemination (AI) is a key breeding technique in the swine industry; however, the lack of reliable biomarkers for semen quality limits its effectiveness. Seminal plasma (SP) contains extracellular vesicles (EVs) that present a promising, non-invasive biomarker for semen quality. This study explores the biochemical profiles of boar SP to assess semen quality through near-infrared spectroscopy (NIRS) and proteomics of SP-EVs. Fresh semen from mature Duroc boars was evaluated based on sperm motility, classifying samples as Passed (≥70%) or Failed (<70%). NIRS analysis identified distinct variations in water structures at specific wavelengths (C1, C5, C12 nm), achieving high accuracy (92.2%), sensitivity (94.2%), and specificity (90.3%) through PCA-LDA. Proteomic analysis of SP-EVs revealed 218 proteins in Passed and 238 in Failed samples. Nexin-1 and seminal plasma protein pB1 were upregulated in Passed samples, while LGALS3BP was downregulated. The functional analysis highlighted pathways associated with single fertilization, filament organization, and glutathione metabolism in Passed samples. Integrating NIRS with SP-EV proteomics provides a robust approach to non-invasive assessment of semen quality. These findings suggest that SP-EVs could serve as effective biosensors for rapid semen quality assessment, enabling better boar semen selection and enhancing AI practices in swine breeding.