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Cervical mucus proteomics approach for swamp buffalo secreted fertility biomarker identification Cover

Cervical mucus proteomics approach for swamp buffalo secreted fertility biomarker identification

Journal of Veterinary Research
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By: ,  ,  ,  ,  ,  ,  ,   and    
Open Access
|May 2026
Figures & tables

Figures & Tables

Fig. 1.

Schematic workflow of cervical-mucus proteomic biomarker discovery in swamp buffalo. LC-MS/MS – liquid chromatography with tandem mass spectrometry; FDR – false detection rate; PCA/PLS-DA – principal component analysis and partial least-squares discriminant analysis; GO – Gene Ontology database; PRM – parallel reaction monitoring

Fig. 2.

Venn diagram of retained features after QC (good swamp-buffalo cervical mucus (GCM) vs poor cervical mucus (PCM))

Fig. 3.

Principal component analysis of swamp-buffalo cervical mucus proteins. (A) Score plot of PC1 vs PC2 showing group separation with 95% confidence ellipses. (B) Biplot illustrating the contribution of selected proteins to group separation. GCM – good cervical mucus; PCM – poor cervical mucus; CLU– clusterin; SPP1 – osteopontin; LTF – lactotransferrin; SLPI – secretory leukocyte protease inhibitor; LCN2 – lipocalin-2; MPO – myeloperoxidase; PTX3 – pentraxin 3; CRP – C-reactive protein; S100A12 – S100 calcium-binding protein A12

Fig. 4.

Volcano plot of differential proteins in good swamp-buffalo cervical mucus and in poor cervical mucus. FC – fold change; MPO – myeloperoxidase; ITGA6 – integrin α6; LTF – lactotransferrin; CLU – clusterin; SLPI – secretory leukocyte protease inhibitor

Fig. 5.

Partial least-squares discriminant analysis variable-importance (VIP) scores for the top 15 proteins in swamp-buffalo cervical mucus. GCM – good cervical mucus; PCM – poor cervical mucus; MPO – myeloperoxidase; LTF – lactotransferrin; LCN2 – lipocalin-2; ITGA6 – integrin α6; CLU – clusterin; ACTA2 – actin α2, smooth muscle; S100A12 – S100 calcium-binding protein A12; SPP1 – osteopontin; PTX3 – pentraxin 3; CRP – C-reactive protein; PRDX6 – peroxiredoxin 6; SLPI – secretory leukocyte protease inhibitor; LGALS3 – galectin-3; SOD1 – superoxide dismutase 1; HSPB6 – heat-shock protein B6

Fig. 6.

Functional enrichment of direction-specific protein sets in swamp-buffalo cervical mucus. GO:MF – Molecular Function in the Gene Ontology database; GO:BP – Biological Process in the Gene Ontology database; GO:CC – Cellular Component in the Gene Ontology database; KEGG – Kyoto Encyclopedia of Genes and Genomes; REAC – Reactome pathway database; WP – WikiPathways; MIRNA – microRNA-related pathways; CORUM – Comprehensive Resource of Mammalian Protein Complexes

Fig. 7.

ROCs for candidate biomarkers in swamp-buffalo cervical mucus. GCM – good cervical mucus; PCM – poor cervical mucus; MPO – myeloperoxidase; LCN2 – lipocalin-2; PTX3 – pentraxin 3; CRP – C-reactive protein; HSPB6 – heat-shock protein B6; SLPI – secretory leukocyte protease inhibitor

Functional annotation of selected proteins and implications for swamp-buffalo cervical mucus quality

SourceTerm IDTerm nameAdjusted P-valueProtein
GO:MFGO:0001849complement component C1q complex binding3.528 × 10-3PTX3; CRP
GO:BPGO:0009620response to fungus1.310 × 10-3MPO; PTX3
GO:BPGO:0044793negative regulation by host of viral process2.456 × 10-2PTX3
GO:CCGO:0005576extracellular region7.656 × 10-7MPO; LTF; LCN2; SPP1; PTX3; CRP; SLPI; LGALS3; HSPB6

Classification performance of biomarker candidates

ProteinDirectionAUC
MPOPCM1
LTFGCM0.86
LCN2GCM1
SPP1GCM0.86
PTX3PCM1
CRPPCM1
SLPIGCM1
LGALS3GCM0.84
HSPB6GCM1

Proteins expressed differentially between good cervical mucus (GCM) and poor cervical mucus (PCM) from swamp buffalo

ProteinFCLog2 (FC)P-valueDirection
MPO1.44 × 10−2-6.1171.31 × 10−13PCM
LTF4.17 × 1015.3812.09 × 10−13GCM
ITGA63.53 × 1028.4622.33 × 10−12GCM
LCN24.632.2113.00 × 10−12GCM
CLU8.94 × 1029.8047.05 × 10−12GCM
S100A121.35 × 10−2-6.2121.19 × 10−11PCM
ACTA25.832.5441.81 × 10−11GCM
SPP11.53 × 1013.9311.88 × 10−11GCM
PTX31.35 × 10−2-6.2114.19 × 10−11PCM
CRP1.34 × 10−2-6.2178.29 × 10−11PCM
PRDX62.61 × 10414.6739.64 × 10−11GCM
ITGB11.54 × 1013.9422.08 × 10−10GCM
SLPI4.26 × 1015.4142.89 × 10−11GCM
LGALS39.35 × 10416.5124.22 × 10−10GCM
SOD12.70 × 10414.7215.82 × 10−10GCM
HSPB67.10 × 1029.4726.99 × 10−10GCM
ANXA52.63 × 10414.6857.67 × 10−11GCM
bTrappin2.151.1079.96 × 10−10GCM
TTR6.99 × 1029.4491.00 × 10−9GCM
VTN4.632.2111.13 × 10−9GCM
HSPB13.43 × 1015.1011.14 × 10−9GCM
FN13.80 × 1028.5691.31 × 10−9GCM
COL3A18.03 × 10416.2931.38 × 10−9GCM
THBS29.60 × 10416.5511.48 × 10−9GCM
GPX12.74 × 10414.7441.61 × 10−9GCM
ACTA19.54 × 10416.5411.87 × 10−9GCM
DOI: https://doi.org/10.2478/jvetres-2026-0025 | Journal eISSN: 2450-8608
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Language: English
Submitted on: Oct 22, 2025
Accepted on: May 8, 2026
Published on: May 14, 2026
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
artificial insemination timing,
decision support,
luminal secretions,
mucosal proteome,
swamp buffalo
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2026 Muhammad Yusuf, Abdul Latief Toleng, Hasrin Hasrin, Abdullah Baharun, Athhar Manabi Diansyah, Ekayanti Mulyati Kaiin, Masturi Masturi, Sahiruddin Sahiruddin, Miftahul Jannah, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution 4.0 License.

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