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Validation of salivary oxytocin and vasopressin as biomarkers in domestic dogs
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Abstract
BACKGROUND: Oxytocin (OT) and Vasopressin (AVP) are phylogenetically conserved neuropeptides with effects on social behavior, cognition and stress responses. Although OT and AVP are most commonly measured in blood, urine and cerebrospinal fluid (CSF), these approaches present an array of challenges including concerns related to the invasiveness of sample collection, the potential for matrix interference in immunoassays, and whether samples can be collected at precise time points to assess event-linked endocrine responses. NEW METHOD: We validated enzyme-linked immunosorbent assays (ELISAs) for the measurement of salivary OT and AVP in domestic dogs. RESULTS: Both OT and AVP were present in dog saliva and detectable by ELISA and high performance liquid chromatography - mass spectrometry (HPLC-MS). OT concentrations in dog saliva were much higher than those typically detected in humans. OT concentrations in the same samples analyzed with and without sample extraction were highly correlated, but this was not true for AVP. ELISA validation studies revealed good accuracy and parallelism, both with and without solid phase extraction. Collection of salivary samples with different synthetic swabs, or following salivary stimulation or the consumption of food led to variance in results. However, samples collected from the same dogs using different techniques tended to be positively correlated. We detected concurrent elevations in salivary and plasma OT during nursing. COMPARISON WITH EXISTING METHODS: There are currently no other validated methods for measuring OT/AVP in dog saliva. CONCLUSIONS: OT and AVP are present in dog saliva, and ELISAs for their detection are methodologically valid.
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“This decision was based on tests for parallelism, spike-recovery, and cross-reactivity/specificity validating the present study’s EIA procedure and suggesting that EIA without extraction may constitute the most accurate assessment of OT in human blood plasma (Carter et al., 2009; MacLean et al., 2017; Lancaster et al., 2017).”
From Associations between oxytocin receptor gene (OXTR) methylation, plasma oxytocin, and attachment across adulthood (2019) · cites this paper as MacLean et al., 2017
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The Association Between Peripheral Oxytocin Levels and Depressive Symptoms in People With HIV
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Salivary Bioscience and Research on Animal Welfare and Conservation Science
Staley, M., & Miller, L. J. (2020) · Springer eBooks · doi:10.1007/978-3-030-35784-9_28 · Cited by 3
AVPR1A distribution in the whole C57BL/6J mouse neonate
Day, K. R., Coleman, A., Greenwood, M. A., & Hammock, E. A. D. (2020) · Scientific Reports · doi:10.1038/s41598-020-71392-1 · Cited by 3
Navarro, N., Jeong, S., Ouassil, N., Mun, J., Leem, E., & Landry, M. P. (2022) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2022.10.05.511026 · Cited by 3
Salivary oxytocin and anxiety in older adults during the COVID-19 pandemic: A cross-sectional study
Kunitake, Y., Imamura, Y., Kunitake, H., Ohishi‐Fukuchi, J., Tateishi, H., Matsushima, J., Murakawa‐Hirachi, T., Kojima, R., Shiraishi, T., Orihashi, R., et al. (2025) · Psychoneuroendocrinology · doi:10.1016/j.psyneuen.2025.107502 · Cited by 4
A Scoping Review on Salivary Oxytocin and Vasopressin Measurement in the Dog
Santis, M. D., Soncin, M., Bertazzo, V., Martelli, L., Fornasiero, D., Mutinelli, F., & Contalbrigo, L. (2025) · Animals · doi:10.3390/ani15162421 · Cited by 2
Strengthening the Foundation of Human–Animal Interaction Research
McCune, S., Kruger, K. A., Griffin, J. A., Esposito, L. E., Bures, R. M., Hurley, K. J., & Gee, N. R. (2019) · Elsevier eBooks · doi:10.1016/b978-0-12-815395-6.00032-8 · Cited by 2
Validation of two immunoassays for oxytocin measurements in human saliva
López-Arjona, M., Cerón, J. M. C., Mateo, S. V., Contreras-Aguilar, M. D., & Martı́nez-Subiela, S. (2024) · PLoS ONE · doi:10.1371/journal.pone.0297539 · Cited by 2
Schroers, M., Juhasz, A., Zablotski, Y., & Meyer‐Lindenberg, A. (2024) · The Veterinary Journal · doi:10.1016/j.tvjl.2024.106148 · Cited by 2
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Gee, N. R., & Wells, D. L. (2024) · Elsevier eBooks · doi:10.1016/b978-0-443-22346-4.00006-8 · Cited by 2
Salivary Oxytocin Has Nonlinear Relationships with Trust and Reciprocity
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Salivary Oxytocin Is Negatively Associated With Religious Faith in Japanese Non-Abrahamic People
Yamada, J., Nakawake, Y., Shou, Q., Nishina, K., Matsunaga, M., & Takagishi, H. (2021) · Frontiers in Psychology · doi:10.3389/fpsyg.2021.705781 · Cited by 3
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RetractedSalivary Biomarkers in Welfare Studies
Escribano, D., & Tecles, F. (2020) · Springer eBooks · doi:10.1007/978-3-030-37681-9_14 · Cited by 1
Metadata
- Authors
- MacLean, E. L., Gesquiere, L. R., Gee, N., Levy, K., Martin, W. L., & Carter, C. S.
- Affiliations
- University of Arizona (US)
- Duke University (US)
- State University of New York (US)
- Canine Companions for Independence (US)
- Indiana University Bloomington (US)
- Corresponding author
- MacLean, Evan L.
- Venue
- Journal of Neuroscience Methods
- Publisher
- Elsevier BV
- Published
- 2018-01
- Volume
- 293
- Pages
- 67-76
- ISSN
- 0165-0270 (ISSN-L)
- OpenAlex
- W2951357445
- Cited by
- 117
- References
- 56
- Citations by year
- 2026: 7 · 2025: 13 · 2024: 17 · 2023: 6 · 2022: 13 · 2021: 19 · 2020: 14 · 2019: 14 · 2018: 11
- FWCI
- 16.48
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Locations
- Journal of Neuroscience Methods (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- UA Campus Repository (The University of Arizona) (acceptedVersion)
- Funders
- Waltham Centre for Pet Nutrition
- Topics
- Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Neuroendocrine regulation and behavior
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Human-Animal Interaction Studies
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Behavior and Welfare Studies
- Keywords
- salivary oxytocin, dogs, biomarkers, ELISA validation, neuropeptides, stress response, saliva collection methods, solid-phase extraction, plasma oxytocin
- MeSH terms
- Animals, Chromatography, High Pressure Liquid, Dogs, Enzyme-Linked Immunosorbent Assay, Female, Lactation, Male, Oxytocin, Saliva, Mass Spectrometry, Vasopressins, Biomarkers
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