Abstract

ObjectiveTo provide an in-depth catalog of the salivary proteome and endogenous peptidome of healthy dogs, evaluate proteins and peptides with antimicrobial properties, and compare the most common salivary proteins and peptides between different breed phylogeny groups.Methods36 healthy dogs without evidence of periodontal disease representing four breed phylogeny groups, based upon single nucleotide polymorphism haplotypes (ancient, herding/sighthound, and two miscellaneous groups). Saliva collected from dogs was pooled by phylogeny group and analyzed using nanoscale liquid chromatography-tandem mass spectrometry. Resulting tandem mass spectra were compared to databases for identification of endogenous peptides and inferred proteins.Results2,491 proteins and endogenous peptides were found in the saliva of healthy dogs with no periodontal disease. All dog phylogeny groups’ saliva was rich in proteins and peptides with antimicrobial functions. The ancient breeds group was distinct in that it contained unique proteins and was missing many proteins and peptides present in the other groups.Conclusions and clinical relevanceUsing a sophisticated nanoscale liquid chromatography-tandem mass spectrometry, we were able to identify 10-fold more salivary proteins than previously reported in dogs. Seven of the top 10 most abundant proteins or peptides serve immune functions and many more with various antimicrobial mechanisms were found. This is the most comprehensive analysis of healthy canine saliva to date, and will provide the groundwork for future studies analyzing salivary proteins and endogenous peptides in disease states.

Highlights

  • Saliva is composed of a complex mixture of enzymes, glycoproteins, immunoglobulins, peptides, inorganic substances, white blood cells, epithelial cells, and microflora, in addition to water

  • A more comprehensive characterization of the salivary proteome and endogenous peptidome of healthy dogs is needed and can provide a valuable groundwork for future studies searching for specific changes in salivary protein composition associated with oral and systemic diseases

  • The dogs were carefully examined by a veterinarian with experience in dental disease to assure the absence of any dental abnormalities, especially periodontal disease

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Summary

Introduction

Saliva is composed of a complex mixture of enzymes, glycoproteins, immunoglobulins, peptides, inorganic substances, white blood cells, epithelial cells, and microflora, in addition to water. For many decades saliva has been considered the reflection of health and disease states of the oral cavity in addition to the whole body [4]. Various mass spectrometry methods are currently available and have been widely used to study the salivary proteins, and smaller, endogenous peptides. A more comprehensive characterization of the salivary proteome and endogenous peptidome of healthy dogs is needed and can provide a valuable groundwork for future studies searching for specific changes in salivary protein composition associated with oral and systemic diseases

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