Clinical use of hormone status determination in canine and feline reproduction

1. Clinical use of hormone status determination in canine and feline reproduction Suppawiwat Ponglowhapan, DVM (Hons), M.S., M.Sc, PhD Department of Obstetrics Gynaecology and Reproduction Faculty of Veterinary Science, Chulalongkorn University

2. Introduction • HormoneHormaein (ภาษากรีก) = Impetus (a moving force, stimulus) แปลว่า กระตุ้น (excite), ก่อกวน (stir up) หรือทำให้เกิดการเปลี่ยนแปลง (to set in motion) • Endocrine The endocrine system is a group of glands that work together and secrete many types of different hormones that regulate the body.

3. Introduction Major endocrine glands. (Male on the left, female on the right.) 1.Pineal gland2.Pituitary gland3.Thyroid gland4.Thymus5.Adrenal gland6.Pancreas7.Ovary8.Testes

4. Introduction • การควบคุมการทำงาน • Autocrine • Paracrine • Endocrine

5. Introduction • Hormone feedback regulatory system • Negative feedback • Positive feedback • Physiological mechanisms of endocrine • Pathologic mechanisms of endocrine • Hormone excess (tumor, exogenous route) • Hormone deficiency (autoimmune, surgery, inflammation) • Hormone resistant (genetic factor)

6. Introduction • สามารถแบ่งฮอร์โมนออกเป็น 5 ชนิด • Amino acid derivatives; dopamine, catecholamines, thyroid hormone • Small neuropeptides; GnRH, TRH, somatostatin, vasopression • Large proteins; LH, FSH, insulin, parathyroid hormone • Steroid hormones; oestrogen, progesterone, testosterone, cortisol • Vitamin derivatives; retinoids

7. Introduction • หน้าที่หลักของฮอร์โมน • Growth and differentiation • Maintenance of homeostasis • Reproduction

8. Introduction • สามารถแบ่งฮอร์โมนออกเป็น 5 ชนิด • Amino acid derivatives; dopamine, catecholamines, thyroid hormone • Small neuropeptides; GnRH, TRH, somatostatin, vasopression • Large proteins; LH, FSH, thyroid hormone, insulin • Steroid hormones; oestrogen, progesterone, testosterone,cortisol • Vitamin derivatives; retinoids

9. Reproduction • Progesterone • Oestrogen • Testosterone • Gonadotrophins: LH & FSH • Thyroid hormone

10. Progesterone Definitions 1. A female hormone and the principal progestational hormone that is made mainly by the corpus luteumin the ovary and by the placenta. Progesterone prepares the lining (endometrium) of the uterus to receive and sustain the fertilized egg and so permits pregnancy. 2. The principal progestational hormone liberated by the corpus luteum, adrenal cortex, and placenta, whose function is to prepare the uterus for the reception and development of the fertilized oocyte by inducing transformation of the endometrium from the proliferative to the secretory stage.

11. Progesterone • A sex steroid hormone • CL, adrenal gland, placenta (cat) • Pregnancy, mammary gland development

12. Progesterone ประโยชน์ของการตรวจโปรเจสเตอโรนในสุนัข • เพื่อกำหนดวันผสม (optimal breeding time) • เพิ่มโอกาสผสมติด • เพิ่มจำนวนลูกต่อครอก • เพื่อตรวจความผิดปกติของการตั้งท้อง • เพื่อทำนายวันคลอด

13. Determination of optimal breeding time in the dog

14. Luteinization of the follicle in the dog

15. Guidelines for interpretation of serum progesterone concentration in the estrous bitch measured by RIA For natural mating or AI using fresh semen, inseminate the day after ovulation For AI using chilled or frozen semen, inseminate 2-4 days after ovulation

16. Predicted time of ovulation based on serum progesterone (n = 50; > 20 breeds) • Key points • Traditional or classical dates for canine breeding: No one-for-all rule • Importance of determining serum progesterone level for canine breeding

17. Determination of optimal breeding time in the dog

18. Determination of optimal breeding time in the dog • LH and ovulation? • Why LH check is less likely to be recommended in practice? • Specificity of LH test • Lab Availability :Thailand?

19. Determination of optimal breeding time in the dog YES • Is there any in-house LH test kit?

20. Determination of optimal breeding time in the dog • Some considerations on the clinical use of LH test kit • Advantages • Price ($230 per box/ 6 tests) • The number of cases • Frequency of the use/ cycle

21. Determination of optimal breeding time in the dog • Unlike other mammals, dogs ovulate when the oestrogen level is declining. • Oestrogen levels can only give us a general idea of when a dog will come into cycle.

22. Vaginal cytology in the cat

23. Determination of optimal breeding time in the dog • Oestrogen and vaginal cytology? basal/ parabasal cell intermediate cell superficial cell

24. Determination of optimal breeding time in the dog • The use of vaginal cytology for canine breeding management • To ensure a dog is in cycle • When to start vaginal swab? • How often? • When to start collecting blood sample for progesterone assay?

25. Recommended workup for determination of optimal breeding time in the bitch • History taking (previous oestrus, first day of vaginal bleeding: Day0) • Physical exam (General and reproductive exam; vaginal discharge, vulvar swelling) • Estrous behaviors • Vaginal smear to confirm the bitch is in estrous stage • Blood collection for progesterone measurement

27. Guidelines for interpretation of serum progesterone concentration in the estrous bitch • Key Notes • Stress and progesterone hypersecretion • Combination of behaviour, vaginal cytology and progesterone • Delivery of live puppies can not be guaranteed.

28. Progesterone ประโยชน์ของการตรวจโปรเจสเตอโรนในสุนัข • เพื่อกำหนดวันผสม (optimal breeding time) • เพิ่มโอกาสผสมติด • เพิ่มจำนวนลูกต่อครอก • เพื่อตรวจความผิดปกติของการตั้งท้อง • เพื่อทำนายวันคลอด

29. Pregnancy monitoring

30. Hypoluteoidism in the bitch • Hypoluteoidism = luteal insufficiency • Progesterone levels > 2.0-2.5 ng/mL are necessary to support pregnancy. • Incidence? • Cause? • Insufficient relaxin low prolactin low progesterone • Endometrial infection/inflammation PGF2 alpha luteolysis

31. Hypoluteoidism in the bitch • Treatment • Medroxyprogesterone acetate (MPA) 0.1 mg/kg PO sid • Altrenogest 0.088 mg/kg PO sid • Progesterone in oil 1-2 mg/kg IM every other day NOTE Discontinued 2-3 days prior to the expected due date

32. Progesterone ประโยชน์ของการตรวจโปรเจสเตอโรนในสุนัข • เพื่อกำหนดวันผสม (optimal breeding time) • เพิ่มโอกาสผสมติด • เพิ่มจำนวนลูกต่อครอก • เพื่อตรวจความผิดปกติของการตั้งท้อง • เพื่อทำนายวันคลอด

33. Prediction of parturition date

34. กำไร • 3 Oct 2009 • Golden Retriever, 2 yrs • 54 days of pregnancy • Severe bloody vaginal discharge • General health status is good • No signs of parturition • X-ray: 3 fetal skeletons (not yet full-term) • U/S: one fetus with slow heat beat (< 180 bpm) • Predicted parturition date by U/S (biparietal diameters) = -7 days

35. กำไร • 6 Oct 2009 • Abortion: one fetus • Progesterone level 1.9 ng/mL • 7 Oct 2009 at night • Deliver 2 living puppies

36. Oestrogen and testosterone • Research purpose • Clinical use is for diagnosis of the presence of gonadal tissue by GnRH stimulating test • Ovarian remnant syndrome oestrogen • Bilateral abdominal cryptorchidism testosterone

37. Ovarian remnant syndrome • How to diagnose wait till the bitch/queen shows oestrus signs GnRH stimulation test • Adverse effects • Owner • Animal

38. Bilateral abdominal cryptorchidism • How to diagnose • Dog: GnRH stimulation test • Cat: penile spine • Adverse effects • Testicular tumour (high incidence) Before castration After castration Testicular tumour

39. Ovarian remnant syndrome • Exner et al., 2008. Theriogenology. Estradiol measurement after GnRH stimulation as a method to diagnose the presence of ovaries in the female domestic cats • GnRH analogue buserelin (Receptal, Intervet) 0.4 microgram/kg IM