1 Clinical use of hormone status determination in canine and feline reproduction Suppawiwat Ponglowhapan, DVM (Hons), M.S., M.Sc, PhDDepartment of Obstetrics Gynaecology and ReproductionFaculty 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)EndocrineThe endocrine system is a group of glands that work together and secrete many types of different hormones that regulate the body.
3 IntroductionMajor 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
10 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.
11 Progesterone A sex steroid hormone CL, adrenal gland, placenta (cat) Pregnancy, mammary gland development
15 Guidelines for interpretation of serum progesterone concentration in the estrous bitch measured by RIASerum Progesterone (ng/mL)Estimated day of ovulationBest breeding daysEstimated parturition from ovulation days+ 2 days+ 4 days (3-6)+ 63 days (62-64)+ 1 days+ 3 days (2-5)0 days+ 2 days (1-4)For natural mating or AI using fresh semen, inseminate the day after ovulationFor 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 pointsTraditional or classical dates for canine breeding: No one-for-all ruleImportance 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 testLab Availability :Thailand?
19 Determination of optimal breeding time in the dog Is there any in-house LH test kit?YES
20 Determination of optimal breeding time in the dog Some considerations on the clinical use of LH test kitAdvantagesPrice ($230 per box/ 6 tests)The number of casesFrequency 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.
23 Determination of optimal breeding time in the dog Oestrogen and vaginal cytology?basal/ parabasal cellintermediate cellsuperficial cell
24 Determination of optimal breeding time in the dog The use of vaginal cytology for canine breeding managementTo ensure a dog is in cycleWhen to start vaginal swab?How often?When to start collecting blood sample for progesterone assay?
25 (previous oestrus, first day of vaginal bleeding: Day0) Physical exam Recommended workup for determination of optimal breeding time in the bitchHistory taking(previous oestrus, first day of vaginal bleeding: Day0)Physical exam(General and reproductive exam; vaginal discharge, vulvar swelling)Estrous behaviorsVaginal smear to confirm the bitch is in estrous stageBlood collection for progesterone measurement
27 Guidelines for interpretation of serum progesterone concentration in the estrous bitch Serum Progesterone (ng/mL)Estimated day of ovulationBest breeding daysEstimated parturition from ovulation days+ 2 days+ 4 days (3-6)+ 63 days (62-64)+ 1 days+ 3 days (2-5)0 days+ 2 days (1-4)Key NotesStress and progesterone hypersecretionCombination of behaviour, vaginal cytology and progesteroneDelivery of live puppies can not be guaranteed.
30 Hypoluteoidism in the bitch Hypoluteoidism = luteal insufficiencyProgesterone levels > ng/mL are necessary to support pregnancy.Incidence?Cause?Insufficient relaxin low prolactin low progesteroneEndometrial infection/inflammation PGF2 alpha luteolysis
31 Hypoluteoidism in the bitch TreatmentMedroxyprogesterone acetate (MPA) 0.1 mg/kg PO sidAltrenogest mg/kg PO sidProgesterone in oil 1-2 mg/kg IM every other dayNOTEDiscontinued 2-3 days prior to the expected due date
34 กำไร 3 Oct 2009 Golden Retriever, 2 yrs 54 days of pregnancy Severe bloody vaginal dischargeGeneral health status is goodNo signs of parturitionX-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 nightDeliver 2 living puppies
36 Oestrogen and testosterone Research purposeClinical use is for diagnosis of the presence of gonadal tissue by GnRH stimulating testOvarian remnant syndrome oestrogenBilateral abdominal cryptorchidism testosterone
37 Ovarian remnant syndrome How to diagnosewait till the bitch/queen shows oestrus signsGnRH stimulation testAdverse effectsOwnerAnimal
38 Bilateral abdominal cryptorchidism How to diagnoseDog: GnRH stimulation testCat: penile spineAdverse effectsTesticular tumour (high incidence)Before castrationAfter castrationTesticular tumour
39 Ovarian remnant syndrome Exner et al., Theriogenology. Estradiol measurement after GnRH stimulation as a method to diagnose the presence of ovaries in the female domestic catsGnRH analogue buserelin (Receptal, Intervet) 0.4 microgram/kg IM