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BMC Vet Res. 2017 Dec 4;13(1):375. doi: 10.1186/s12917-017-1296-9.

Influence of different sample preparation strategies on the proteomic identification of stress biomarkers in porcine saliva.

Author information

1
Department of Animal Medicine and Surgery, Regional Campus of International Excellence "Campus Mare Nostrum", Hospital Veterinario 4 planta, University of Murcia, 30100, Espinardo, Murcia, Spain.
2
VetCore Facility for Research, University of Veterinary Medicine Vienna, Veterinaerplatz 1, A-1210, Vienna, Austria.
3
Department of Animal Medicine and Surgery, Regional Campus of International Excellence "Campus Mare Nostrum", Hospital Veterinario 4 planta, University of Murcia, 30100, Espinardo, Murcia, Spain. ftecles@um.es.

Abstract

BACKGROUND:

The influence of two different sample treatments comprising the enrichment of glycoproteins by boronic acid and dynamic range compression by hexapeptide libraries, on the detection of stress markers in saliva of pigs was evaluated in this study. For this purpose, saliva samples collected before and after the application of an acute stress model consisting of nasal restraining in pigs were processed without any treatment and with the two different treatments mentioned above. Protein separation by two-dimensional gel electrophoresis (2-DE) followed by identification of proteins using MALDI-TOF/TOF mass spectrometry (MS) was used as proteomic technique.

RESULTS:

The application of each of the two different sample treatment protocols allowed the identification of unique proteins that could be potential salivary acute stress markers in pigs: lipocalin 1, protein S100-A8 and immunoglobulin M by enrichment of glycoproteins; protein S100-A9, double headed protease inhibitor submandibular gland, and haemoglobin by dynamic range compression; and protein S100-A12 by both protocols. Salivary lipocalin, prolactin inducible protein, light chain of immunoglobulins, adenosine deaminase and carbonic anhydrase VI were identified as potential markers in untreated saliva as well as one of the other treatments.

CONCLUSION:

The use of different procedures allowed the detection of different potential stress markers. Although from a practical point of view, the use of saliva without further treatment as well as the enrichment of glycoproteins are less expensive and easy to do procedures.

KEYWORDS:

Glycoprotein-enrichment; Hexapeptide libraries; Pig; Proteomics; Saliva

PMID:
29202764
PMCID:
PMC5716369
DOI:
10.1186/s12917-017-1296-9
[Indexed for MEDLINE]
Free PMC Article

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