Cervical mucus proteomics approach for swamp buffalo secreted fertility biomarker identification

Abstract

Introduction: ) remains constrained by the lack of molecular tools to identify fertile females before service. The aim of this study was to discover biologically grounded biomarkers of fertility. Material and Methods: Cervical mucus was profiled using high-resolution proteomic analysis, with quantitative and functional enrichment used to prioritise biologically relevant fertility markers. Results: The cervical-mucus proteome exhibited a conserved secreted core with marked quantitative shifts that separated favourable-quality from poor-quality mucus. Favourable mucus was characterised by a barrier- and adhesion-associated profile with antimicrobial and antioxidant features, its proteins including integrin/extracellular matrix interactors, redox enzymes, lipocalin 2 (LCN2), secretory leukocyte protease inhibitor (SLPI) and heat-shock protein B6 (HSPB6). Poor mucus showed a neutrophil-acute-phase axis with complement engagement and oxidative chemistry, typified by myeloperoxidase (MPO), pentraxin 3 (PTX3) and C-reactive protein (CRP). Functional enrichment emphasised extracellular and host-defence processes consistent with these opposing mucosal states. These six proteins were consistently the leading discriminants and had coherent biological roles. Conclusion: Swamp-buffalo cervical mucus is variously in a barrier-competent state and a neutrophil-dominant state. This dichotomy provides a mechanistic explanation for fertility differences at the cervix and yields actionable, secreted protein biomarkers - LCN2/SLPI/HSPB6 in favourable mucus and MPO/PTX3/CRP in poor mucus - for targeted assays and pre-service screening in tropical production systems.

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