Clinical use of hormone status determination in canine

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

Introduction • Hormone Hormaein (ภาษากรก ) = Impetus (a moving force, stimulus) แปลวา กระตน (excite), กอกวน (stir up) หรอทำใหเกดการเปลยนแปลง (to set in motion) • Endocrine The endocrine system is a group of glands that work together and secrete many types

Introduction Major endocrine glands. (Male on the left, female on the right. ) 1. Pineal gland 2. Pituitary gland 3. Thyroid gland 4. Thymus 5. Adrenal gland 6. Pancreas 7. Ovary 8. Testes

Introduction • การควบคมการทำงาน ▫ Autocrine ▫ Paracrine ▫ Endocrine

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)

Introduction • สามารถแบงฮอรโมนออกเปน 5 ชนด 1. Amino acid derivatives; dopamine, catecholamines, thyroid hormone 2. Small neuropeptides; Gn. RH, TRH, somatostatin, vasopression 3. Large proteins; LH, FSH, insulin, parathyroid hormone 4. Steroid hormones; oestrogen, progesterone, testosterone, cortisol

Introduction • หนาทหลกของฮอรโมน 1. Growth and differentiation 2. Maintenance of homeostasis 3. Reproduction

Introduction • สามารถแบงฮอรโมนออกเปน 5 ชนด 1. Amino acid derivatives; dopamine, catecholamines, thyroid hormone 2. Small neuropeptides; Gn. RH, TRH, somatostatin, vasopression 3. Large proteins; LH, FSH, thyroid hormone, insulin 4. Steroid hormones; oestrogen, progesterone, testosterone, cortisol 5. Vitamin derivatives; retinoids

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

Progesterone Definitions 1. A female hormone and the principal progestational hormone that is made mainly by the corpus luteum in 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

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

Progesterone ประโยชนของการตรวจโปรเจสเตอโรนในสนข 1. เพอกำหนดวนผสม (optimal breeding time) เพมโอกาสผสมตด เพมจำนวนลกตอครอก 2. เพอตรวจความผดปกตของการตงทอง 3. เพอทำนายวนคลอด

Determination of optimal breeding time in the dog

Luteinization of the follicle in the dog

Guidelines for interpretation of serum progesterone concentration in the estrous bitch measured by RIA Serum Progesterone (ng/m. L) Estimated day of ovulation Best breeding days Estimated parturition from ovulation days 1. 0 -1. 9 + 2 days + 4 days (3 -6) + 63 days (62 -64) 2. 0 -3. 9 + 1 days + 3 days (2 -5) + 63 days (62 -64) 4. 0 -10. 0 0 days + 2 days (1 -4) + 63 days (62 -64) 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

Predicted time of ovulation based on serum progesterone (n = 50; > 20 breeds) Key points 1. Traditional or classical dates for canine breeding: No one-for-all rule 2. Importance of determining serum progesterone level for canine breeding

Determination of optimal breeding time in the dog

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?

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

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

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.

Vaginal cytology in the cat

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

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?

Recommended workup for determination of optimal breeding time in the bitch • History taking (previous oestrus, first day of vaginal bleeding: Day 0) • Physical exam (General and reproductive exam; vaginal discharge, vulvar swelling) • Estrous behaviors • Vaginal smear to confirm the bitch is in estrous stage

Guidelines for interpretation of serum progesterone concentration in the estrous bitch Serum Progesterone (ng/m. L) Estimated day of ovulation Best breeding days Estimated parturition from ovulation days 1. 0 -1. 9 + 2 days + 4 days (3 -6) + 63 days (62 -64) 2. 0 -3. 9 + 1 days + 3 days (2 -5) + 63 days (62 -64) 4. 0 -10. 0 0 days + 2 days (1 -4) + 63 days (62 -64) Key Notes 1. Stress and progesterone hypersecretion 2. Combination of behaviour, vaginal cytology and progesterone 3. Delivery of live puppies can not be guaranteed.

Progesterone ประโยชนของการตรวจโปรเจสเตอโรนในสนข 1. เพอกำหนดวนผสม (optimal breeding time) เพมโอกาสผสมตด เพมจำนวนลกตอครอก 2. เพอตรวจความผดปกตของการตงทอ ง 3. เพอทำนายวนคลอด

Pregnancy monitoring

Hypoluteoidism in the bitch • Hypoluteoidism = luteal insufficiency • Progesterone levels > 2. 0 -2. 5 ng/m. L are necessary to support pregnancy. • Incidence? • Cause? ▫ Insufficient relaxin low prolactin progesterone ▫ Endometrial infection/inflammation alpha luteolysis low PGF 2

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

Progesterone ประโยชนของการตรวจโปรเจสเตอโรนในสนข 1. เพอกำหนดวนผสม (optimal breeding time) เพมโอกาสผสมตด เพมจำนวนลกตอครอก 2. เพอตรวจความผดปกตของการตงทอง 3. เพอทำนายวนคลอด

Prediction of parturition date

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)

กำไร • 6 Oct 2009 • Abortion: one fetus • Progesterone level 1. 9 ng/m. L • 7 Oct 2009 at night • Deliver 2 living puppies

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

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

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

Ovarian remnant syndrome • Exner et al. , 2008. Theriogenology. Estradiol measurement after Gn. RH stimulation as a method to diagnose the presence of ovaries in the female domestic cats • Gn. RH analogue buserelin (Receptal, Intervet) 0. 4 microgram/kg IM