Pituitary Pars Intermedia Dysfunction (PPID) remains the most commonly diagnosed endocrinopathy in the horse. Dopaminergic neurons in the hypothalamus degenerate leading to a loss of the normal tonic inhibition of the pars intermedia. This leads to overproduction of many peptides including pro-opio melanocortin, α-melanocyte-stimulating hormone, corticotrophin-like intermediate peptide and adrenocorticotropic hormone (ACTH). Clinical signs of PPID include hirsutism, sweating, abnormal fat deposition, muscle wastage, laminitis, lethargy and polydipsia.

Diagnosis in advanced cases is often easy based on the presence of clinical signs. However, laboratory diagnosis is important for less advanced cases, and to aid in monitoring the response to treatment. Understanding the limitations of the currently available diagnostic tests is also important to avoid overdiagnosis of the disease and unnecessary treatment.

An excellent resource is the Equine Endocrinology Group website.

Recommended first line test:

Basal plasma ACTH concentration

Sampling protocol:

  • Collect a single EDTA blood sample at any time of day
  • Chill within 3 hours
  • Separate the plasma by centrifugation or gravity
  • Ship to the laboratory overnight with cool packs (maximum delay 48 hours)
  • If there will be a delay between sample collection and analysis then centrifuged plasma can be frozen and is then stable for many weeks. Gravity separated plasma should not be frozen as this will lead to a spurious increase in ACTH concentration

Test Interpretation

This is the recommended first line test for older horses in which you have a strong suspicion of clinical disease based on the presence of clinical signs.

The test should be avoided in young horses (less than 10 years old), or those without any clinical signs, as the positive predictive power of the test in these circumstances is low and many false positives occur. Horses with early disease often have normal or equivocal ACTH concentrations, and further testing with a TRH stimulation test, or repeated monitoring may be needed to evaluate further.

Season has a major impact on ACTH concentration in both normal and PPID affected horses and it is important that results are interpreted in line with seasonal reference ranges.

As there is significant variation in ACTH concentrations between individuals, a single positive threshold is not recommended and an equivocal range is reported. Results that fall in this range should be interpreted in conjunction with the clinical signs and age of the horse. Depending on these factors it may be appropriate to perform a TRH stimulation test, monitor and re-test or treat the horse.

The following table summarises the suggested cut-offs using the Tosoh AIA-900. Results are listed in pg/ml.

Time of Year (Northern Hemisphere)PPID UnlikelyEquivocal (requires strong clinical signs, retesting or TRH stimulation test to confirm diagnosis)PPID Likely
December – June<1212-32>32
July and November<1616-44>44
August<2222-52>52
September – October<2323-58>58

The breed of horse being tested should also be considered. Pony breeds, Arabians and donkeys have all been shown to have higher plasma ACTH concentrations especially in the Autumn months. Concurrent disease, stress and travel can also have an impact on ACTH concentration and the presence or absence of these factors should be considered when interpreting any result.

Thyrotropin releasing hormone stimulation test (TRHST)

This is currently the recommended dynamic test for the diagnosis of PPID. The test relies on an excessive pituitary response to the administration of thyrotropin-releasing hormone (TRH) in horses with PPID when compared to normal horses.  However, the test still has limitations and ongoing validation and research is needed to help us fully understand how to interpret the results. Usefulness of the test is limited to the period from January to June and updated diagnostic thresholds will be published once established for the Tosoh AIA-900 analyser.

Sampling protocol

  • Horse may have normal access to hay prior to the test but should not have any concentrate feed for 12 hours before testing.
  • Collect an EDTA sample for baseline measurement of ACTH.
  • Inject 1mg TRH intravenously in horses over 250kg. Use 0.5mg intravenously in horses or ponies less than 250kg.
  • Collect a second EDTA sample exactly 10 minutes later.
  • The plasma should be handled as described in the ACTH section.

Interpreting the result

The following table summarises the current diagnostic thresholds using the Immulite analyser.  New thresholds are being generated for the Tosoh AIA-900 analyser.

Time of YearNegativeEquivocal (requires strong clinical signs, retesting or TRH stimulation test to confirm diagnosis)PPID Likely
January to June<100100 – 200>200
July to December<100TRH stimulation testing can only be used to identify negative cases in these months due to many false positives

Availability of TRH

Compounded TRH can be purchased here

The product is sold as Thyrotropic Releasing Hormone (TRH) 1mg/5ml

Side effects of TRH administration

Side effects are rare but include trembling, lip-smacking and flehmen type behaviour.

Other tests in horses with PPID

Insulin dysregulation is a common concurrent problem in horses with PPID. It is good practice to evaluate this as a minimum by measurement of resting insulin and glucose concentrations or by performing a dynamic sugar challenge test. This can be useful when deciding about the need for pergolide treatment in a horse with equivocal results. The presence of hyperinsulinaemia may be associated with an increased risk of laminitis and consequently a greater need for treatment for PPID.

Further information

ACTH and Insulin in Equids – the significance of the analytical method.

 

References:

  1. Adams A. Evaluating seasonal influences on hormone responses to a diagnostic test advocated for early diagnosis of pituitary pars intermedia dysfunction. 2017.Havemeyer International Endocrine Symposium, Miami
  2. Beech J, Boston R, Lindborg S, Russell GE. Adrenocorticotropin concentration following administration of thyrotropin-releasing hormone in healthy horses and those with pituitary pars intermedia dysfunction and pituitary gland hyperplasia. J Am Vet Med Assoc. 2007; 231:417-26
  3. Beech J, Boston R, Lindborg S. Comparison of Cortisol and ACTH Responses after Administration of Thyrotropin Releasing Hormone in Normal Horses and Those with Pituitary Pars Intermedia Dysfunction. J Vet Intern Med. 2011; 25:1431-8
  4. Copas VE, Durham AE. (2012) Circannual variation in plasma adrenocorticotropic hormone concentrations in the UK in normal horses and ponies, and those with pituitary pars intermedia dysfunction. Equine Vet J. 2012 Jul;44(4):440–3
  5. Diez de Castro E, Lopez I, Cortes B, Pineda C, Garfia B, Aguilera-Tejero E. Influence of feeding status, time of the day, and season on baseline adrenocorticotropic hormone and the response to thyrotropin releasing hormone-stimulation test in healthy horses. Domest Anim Endocrinol. 2014 Jul;48:77-83.
  6. Durham AE, Clarke BR, Potier JF, Hammarstrand RD, Malone GL (2020) Clinically and temporally specific diagnostic thresholds for plasma ACTH in the horse Equine Vet J May 29.
  7. Funk RA, Stewart AJ, Wooldridge AA, Kwessi E, Kemppainen RJ, Behrend EN, Zhong Q, Johnson AK. Seasonal changes in plasma adrenocorticotropic hormone and α-melanocyte-stimulating hormone in response to thyrotropin-releasing hormone in normal, aged horses. J Vet Intern Med. 2011 May-Jun;25(3):579-85.
  8. Goodale L, Frank N, Hermida P, D’Oench S. Evaluation of a thyrotropin-releasing hormone solution stored at room temperature for pituitary pars intermedia dysfunction testing in horses. Am J Vet Res. 2015; 76(5):437-44
  9. Kirkwood, N. C., Hughes, K. J., & Stewart, A. J. (2022). Pituitary Pars Intermedia Dysfunction (PPID) in Horses. Veterinary Sciences, 9(10), 556.
  10. McFarlane D, Maxwell LK. Establishment of a reference interval for plasma ACTH concentration in aged horses. 2017. Havemeyer International Endocrine Symposium, Miami
  11. McFarlane D. Diagnostic Testing for Equine Endocrine Diseases: Confirmation Versus Confusion. Vet Clin North Am Equine Pract. 2019 Aug;35(2):327-338
  12. Rendle DR, Laboratory diagnosis of the endocrine causes of laminitis Livestock July/August 2017, Volume 22 No 4
  13. Restifo MM, Frank N, Hermida P, Sanchez-Londoño A. Effects of withholding feed on thyrotropin-releasing hormone stimulation test results and effects of combined testing on oral sugar and thyrotropin-releasing hormone stimulation test results in horses. Am J Vet Res. 2016; 77:738-48
  14. Stewart, A. J., Ireland, J. L., Durham, A. E., & McGowan, C. M. (2023). Diagnosis of equine pituitary pars intermedia dysfunction. Veterinary journal (London, England : 1997), 300-302, 106036.