engleski

Biomarkers for diagnosing the retained placenta in cows-does the analysis of serum proteome keep the secret?

Question: Retained placenta (RP) is a common disorder during the puerperium in dairy cows with a high negative impact on the health condition of the cows and the milk production. The initiation of the RP therapy when the clinical signs are already present has low effectiveness, which highlights the necessity for the biomarkers that would indicate the risk for the RP before the occurrence of the signs. The study aimed to investigate the serum proteome as the source of the biomarkers for diagnosing the RP. Methods: The study included nine cows with the RP and six with the physiologic puerperium. The serum proteins were reduced, alkylated, precipitated with acetone, digested with trypsin, and labeled with the tandem mass tags. The liquid chromatography-tandem mass spectrometry platform consisted of the Ultimate 3000 RSLCnano system (Dionex, Germering, Germany) coupled to a Q Exactive Plus mass spectrometer (Thermo Fisher Scientific, Bremen, Germany). The peptides were separated using the linear gradient and scanned in the positive ion mode using the data- dependent analysis Top8 method. For the identification and relative quantitation following the SEQUEST algorithm, the Proteome Discoverer® software was employed. Further, the data were analyzed with the R statistical package and Protein Analysis Through Evolutionary Relationship (PANTHER) classification tool. Results: In total, 651 proteins were quantified. Fiftyfour had different relative abundance in the cows with RP and those with the physiological puerperium, whereby approximately two-thirds were more abundant in the RP cases. The main classes containing the proteins with the different abundances were the modulators of protein-binding activity and the protein modifying enzymes. The gene ontology results indicated that these proteins posse catalytic activity, binding, and regulatory functions in eleven biological processes. The highest increase in abundance was for haptoglobin, followed by lipopolysaccharide- binding protein, inter-alpha-trypsin inhibitor heavy chain, and glutathione peroxidase. The cows with RP had the highest decrease in apolipoprotein A-IV, while it was less protruding for C-type lectin domain family 3 member B, tetranectin, and serpin family D member 1. Conclusion: The serum proteome analysis deciphered several aspects of the RP molecular pathology. Also, it identified several proteins with the potential to serve as the RP biomarkers after further clinical validation.

retained placenta, cows, proteomics

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