Surreal illustration of a horse silhouette with saliva droplets, representing pain detection.

Is Your Horse in Pain? A New Saliva Test Could Offer Answers

Kai Mendoza in Health & Wellness December 2025 • 4 min read.

"Researchers explore how salivary alpha-amylase and cortisol levels can help detect acute abdominal pain in horses, providing a non-invasive way to assess their well-being."

Recognizing pain in horses can be challenging, especially when dealing with acute abdominal issues like colic. Traditional methods often rely on subjective assessments and invasive procedures, creating a need for more objective and stress-free evaluation techniques.

A recent study has explored the potential of salivary alpha-amylase (sAA) and salivary cortisol as biomarkers for pain and stress in horses experiencing acute abdominal disease. These substances, easily collected through saliva, could provide valuable insights into a horse's condition without the need for blood samples.

This article delves into the findings of this research, explaining how sAA and cortisol levels in saliva can indicate pain-induced stress in horses. We'll explore the study's methodology, key results, and the implications for improved equine welfare.

Decoding Saliva: How the Study Measured Pain in Horses

Surreal illustration of a horse silhouette with saliva droplets, representing pain detection.

The study, published in BMC Veterinary Research, involved analyzing saliva samples from 19 horses diagnosed with acute abdominal disease and comparing them to a control group of 11 healthy horses. The goal was to determine if there were significant differences in sAA and cortisol levels between the two groups.

Researchers collected saliva samples before any potentially painful procedures were performed. They used the Equine Acute Abdominal Pain Scale (EAAPS-1) to assess the severity of pain based on observed behaviors. Blood samples were also taken to measure other indicators like plasma lactate and serum amyloid A (SAA).

Salivary alpha-amylase (sAA) activity: Measured using a colorimetric commercial kit, following established scientific methods.
Salivary cortisol levels: Analyzed with an immunoassay system, ensuring accurate and reliable measurements.
Statistical analysis: Employed to compare the data between the two groups and identify any significant correlations between biomarker levels and pain scores.

The stability of sAA and salivary cortisol in saliva samples was tested by storing samples at room temperature (25 °C) and in refrigeration (4 °C) for 48 h. Measurements were taken just after saliva collection, 24 h and 48 h later.

Saliva: A Promising Tool for Assessing Equine Pain

The study's findings revealed that horses with acute abdominal disease had significantly higher sAA and cortisol levels in their saliva compared to healthy horses. Importantly, sAA activity showed a strong correlation with the EAAPS-1 pain scale, indicating its potential as a reliable indicator of pain.

While salivary cortisol levels were elevated in horses with colic, they did not correlate as strongly with the pain scale. This suggests that sAA may be a more sensitive marker of pain-induced stress in these cases.

These preliminary results pave the way for further research into the use of saliva as a non-invasive diagnostic tool for assessing pain and stress in horses. By understanding the relationship between salivary biomarkers and equine health, veterinarians can improve animal welfare and provide more effective treatment strategies.

About this Article -

This article was crafted using a human-AI hybrid and collaborative approach. AI assisted our team with initial drafting, research insights, identifying key questions, and image generation. Our human editors guided topic selection, defined the angle, structured the content, ensured factual accuracy and relevance, refined the tone, and conducted thorough editing to deliver helpful, high-quality information.See our About page for more information.

This article is based on research published under:

DOI-LINK: 10.1186/s12917-018-1482-4, Alternate LINK

Title: Salivary Alpha-Amylase Activity And Cortisol In Horses With Acute Abdominal Disease: A Pilot Study

Subject: General Veterinary

Journal: BMC Veterinary Research

Publisher: Springer Science and Business Media LLC

Authors: María Dolores Contreras-Aguilar, Damián Escribano, María Martín-Cuervo, Fernando Tecles, Jose Joaquín Cerón

Published: 2018-05-10

Everything You Need To Know

How did the researchers measure pain in horses using saliva?

The study published in BMC Veterinary Research, measured salivary alpha-amylase (sAA) and salivary cortisol levels in horses experiencing acute abdominal issues, comparing them to a control group. The researchers collected saliva samples before any painful procedures, used the Equine Acute Abdominal Pain Scale (EAAPS-1) to assess pain, and analyzed the samples to determine differences in sAA and cortisol levels between the groups.

What specific methods were used to analyze salivary alpha-amylase and salivary cortisol levels?

Salivary alpha-amylase (sAA) activity was measured using a colorimetric commercial kit, following scientific methods. Salivary cortisol levels were analyzed with an immunoassay system for accurate measurements. Statistical analysis was used to compare data between the groups and identify correlations between biomarker levels and pain scores. Additionally, stability tests were conducted by storing samples at room temperature (25 °C) and in refrigeration (4 °C) for 48 hours to ensure reliability.

What were the key findings of the study regarding salivary alpha-amylase, salivary cortisol, and their relation to pain?

The study found that horses with acute abdominal disease had significantly higher salivary alpha-amylase (sAA) and salivary cortisol levels compared to healthy horses. The sAA activity also strongly correlated with the Equine Acute Abdominal Pain Scale (EAAPS-1), indicating its potential as a reliable indicator of pain. These findings suggest that saliva samples can provide valuable insights into a horse's condition without invasive procedures.

What are the potential benefits of using salivary alpha-amylase and salivary cortisol measurements for assessing equine pain?

Measuring salivary alpha-amylase (sAA) and salivary cortisol levels offers a non-invasive method to assess a horse's well-being, reducing the stress associated with traditional blood tests. This is particularly beneficial for horses with acute abdominal issues like colic, where recognizing pain can be challenging. By using saliva samples, veterinarians can gain objective insights into the horse's condition, potentially leading to earlier and more effective pain management.

Beyond salivary alpha-amylase and salivary cortisol, what other areas could future research explore to improve pain assessment in horses?

While the study focuses on salivary alpha-amylase (sAA) and salivary cortisol, future research could explore other potential salivary biomarkers to enhance diagnostic accuracy. Additional factors such as inflammatory markers or specific proteins related to abdominal pain could provide a more comprehensive understanding of a horse's condition. Furthermore, longitudinal studies tracking biomarker levels over time could help in monitoring treatment response and predicting outcomes in horses with acute abdominal disease.