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
Introduction
• Hormone  Hormaein (ภาษากรี ก)
= Impetus (a moving force, stimulus)
แปลว่า กระตุ้น (excite), ก่อกวน (stir up) หรื อทาให้ เกิดการเปลี่ยนแปลง
(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.
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
• การควบคุมการทางาน
▫ 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)
▫ Hormone resistant (genetic factor)
Introduction
• สามารถแบ่งฮอร์ โมนออกเป็ น 5 ชนิด
1. Amino acid derivatives; dopamine, catecholamines, thyroid
hormone
2. Small neuropeptides; GnRH, TRH, somatostatin, vasopression
3. Large proteins; LH, FSH, insulin, parathyroid hormone
4. Steroid hormones; oestrogen, progesterone, testosterone, cortisol
5. Vitamin derivatives; retinoids
Introduction
• หน้ าที่หลักของฮอร์ โมน
1. Growth and differentiation
2. Maintenance of homeostasis
3. Reproduction
Introduction
• สามารถแบ่งฮอร์ โมนออกเป็ น 5 ชนิด
1.
2.
3.
4.
5.
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
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 to the secretory stage.
Progesterone
• A sex steroid hormone
• CL, adrenal gland, placenta (cat)
• Pregnancy, mammary gland development
Progesterone
ประโยชน์ของการตรวจโปรเจสเตอโรนในสุนขั
1.เพื่อกาหนดวันผสม (optimal breeding time)


เพิ่มโอกาสผสมติด
เพิ่มจานวนลูกต่อครอก
2.เพื่อตรวจความผิดปกติของการตังท้
้ อง
3.เพื่อทานายวันคลอด
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/mL)
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
• Is there any in-house LH test kit? YES
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: 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
Guidelines for interpretation of serum progesterone
concentration in the estrous bitch
Serum Progesterone
(ng/mL)
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
ประโยชน์ของการตรวจโปรเจสเตอโรนในสุนขั
1.เพื่อกาหนดวันผสม (optimal breeding time)


เพิ่มโอกาสผสมติด
เพิ่มจานวนลูกต่อครอก
2.เพื่อตรวจความผิดปกติของการตัง้ ท้ อง
3.เพื่อทานายวันคลอด
Pregnancy monitoring
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
▫ Endometrial infection/inflammation
low progesterone
PGF2 alpha
luteolysis
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
ประโยชน์ของการตรวจโปรเจสเตอโรนในสุนขั
1.เพื่อกาหนดวันผสม (optimal breeding time)


เพิ่มโอกาสผสมติด
เพิ่มจานวนลูกต่อครอก
2.เพื่อตรวจความผิดปกติของการตังท้
้ อง
3.เพื่อทานายวันคลอด
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)
▫ Predicted parturition date by U/S (biparietal diameters) = -7 days
กำไร
• 6 Oct 2009
• Abortion: one fetus
• Progesterone level 1.9 ng/mL
• 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 GnRH
stimulating test
 Ovarian remnant syndrome
oestrogen
 Bilateral abdominal cryptorchidism
testosterone
Ovarian remnant syndrome
• How to diagnose
wait till the bitch/queen shows oestrus signs
GnRH stimulation test
• Adverse effects
▫ Owner
▫ Animal
Bilateral abdominal cryptorchidism
• How to diagnose
▫ Dog: GnRH stimulation test
▫ Cat: penile spine
• Adverse effects
Before castration
After castration
▫ Testicular tumour (high incidence)
Testicular tumour
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