Breeding Management and Assisted Reproduction

1. Breeding Management and Assisted Reproduction Dr. N. Matthew Ellinwood, D.V.M., Ph.D. February 2, 2012 Iowa State University College of Agriculture and Life Sciences

2. Breeding Management • How soon, i.e. what age • How often • Breeding techniques

3. Introduction

4. Bitch Review • First Cycle • 6 to 18 months of age • Proestrus • Vaginal discharge to standing heat • Estrus • Standing heat to refusal • Diestrus • Refusal of breeding to whelping • Anestrus • Whelping to onset of next cycle

5. Endocrinology of the CycleReview • Proestrus • Estrogen, produced by follicles • Growth and bleeding in the uterus • Cellular thickening of vaginal canal • Estrus • Estrogen is decreasing, while progesterone is increasing • Standing heat, 48 hours post LH peak (ovulation) • ~95% superficial cells • Diestrus • Pregnancy/pseudopregnancy, progesterone dominant hormone • Behavior: the bitch refuses to stand for breeding • Cytology: decline in % of superficial cells on vaginal cytology • Most reliable non-hormonal milestone of cycle

6. Other clues to onset of estrus • Not 100 % reliable • Vulva may become less swollen and/or turgid • Bloody vaginal discharge may change to a straw colored discharge • While these can help, the most reliable method will remain monitoring vaginal cytology and behavior changes in the bitch and the male

7. Overview of the cycle http://arbl.cvmbs.colostate.edu/hbooks/pathphys/reprod/vc/

8. First Breedings • > 2 years of age for stud and bitch • Hip certification and other health screenings • Bitch • No more often than every other cycle • Every two years recommended • Stud • Avoid overuse • Population • Progeny evaluation

9. Primary cause of “infertility” • Most bitches who fail to conceive have normal fertility • Most males who fail to settle a bitch with pups have normal fertility • The single greatest cause of infertility is failure to breed at the right time

10. Tools and concepts necessary for successful breedings • The average bitch does not exist • Trusting to “chemistry” . . . How to assess normal courting and mating behavior • Vaginal cytology . . . The tried and true method in simple breeding management • The power of the pen . . . Keeping records for the lifetime of your bitch

11. Averages • Most bitches will follow a certain pattern in their cycle through the middle years of their reproductive career • Keeping records • Start and end of vaginal discharge • Changes in behavior and cytology results • Breeding and whelping dates

12. Chemistry • The guys • Lack of, increase of, and loss of ardor • The girls • La donna è mobile • The old maid • The social misfit • Atmosphere • Territory, environment, security

13. Vaginal cytology • Parabasal cells • Intermediate cells • Superficial cells

15. Pre-breeding routine medical concerns • When to pursue them, who to contact • Physical exams • Reproductive exams • Laboratory assessments • Cultures? • Serologies? • Brucella canis

16. Things to have in order • Veterinary contacts • Secure and appropriate facility for managing the breeding • Breeding only area

17. Courtship and Mating • Due no harm . . . ensuring the safety of the animals • Hand breeding • Attended breedings

19. Breeding plan • Assumes no travel • Both bitch and stud on site • +/- cytology • No first timers • No breeds that may warrant AI

20. Breeding on the next heat • Four months (or normal cycle -2 months) after onset of last heat • Pursue medical check up • Ensure bitch is not over- or under-conditioned • Begin weekly vulvar exams (or how you can miss the obvious) • No intact male at home • Long or dark haired bitch • Fastidious bitch • Very mild changes in the external tract

21. Onset of the estrous cycle • First day discharge - baseline cytology if possible • Monitor every 3 to 4 days until 90% superficial cells • Or • Monitor daily for standing estrus with an intact proven male • Confirm with cytology if possible

22. Breeding schedule • Breed every other day for the first two or three breedings • After initial breedings • Test behavior daily • Breed every 3 to 4 days until diestrus shift, or until bitch refuses or male loses interest • Corroborate with a cytology if possible

23. Breeding DogsWhat to Expect • Mounting, intromission and thrusting • Short (1-5 minutes) • Tie • Dismounting, retroflexion • Long 5-45 minutes • Health and safety concerns

24. Planning for Parturition • Diestrus shift • Cytology or behavior • Day 9 post LH peak • Date of parturition • Diestrus shift + 56 days (+/- 1 day)

25. Behavioral complications • Older maiden bitches • Under socialized bitches • Aggressive or uninterested stud • Must confirm status within estrous cycle by cytology • If poor correspondence between behavior and cycle, pursue artificial insemination • Continue to test for onset of standing estrus

26. Breeding Cats • Plan for in house cattery • Mature proven breeding stock • No infectious diseases

27. Queen • > 1 year of age • No more often than one litter per year • Secure breeding cage or run • Not the queen’s cage • Some shelter within breeding enclosure • One tom per breeding run • Up to a dozen queens per breeding run

28. Timed versus Un-timed • First palpation best method to estimate breeding and hence parturition dates • Un-timed breeding • Co-house queen and tom until pregnancy achieved • Weekly palpation • Begin in February/March • Mating behaviors may make breedings obvious • Mounting, neck biting, screaming, post-coital behavior • Timed breeding • Queen to breeding run 2 days/week • Same two days each week • Palpate thrice weekly

29. Estimating Parturition • Timed matings example • Breed from Monday morning to Tuesday evening • Palpate on Thursday (day 18), Saturday (day 20), and Monday (day 22), beginning after third week of breedings • Calculate back to week of ovulation after palpated positive • 66 days +/- 1 day from ovulation • 66 days -1 to +2 days from Monday bred • Expect kittens Tuesday – Friday (9 weeks, 3 days) • Most likely Wednesday to Thursday

30. Review • Canine • Management to ensure safety, security, paternity, fertility, and best estimate of parturition • Diestrus • Behavior • Cytology • Feline • Timed versus un-timed pregnancies • Palpations

31. Tools for Managing Successful Canine Fertility

32. Averages versus Reality • Know the average length of the various parts of the estrus cycle • Know how they can deviate

33. Behavior • Know how to assess behavior • Flagging, riding, etc.

34. Flagging

35. Record Keeping

36. Vaginal Cytology

37. Observed or Hand Breeding

38. Vulvar Swelling of Proestrus/Estrus

39. Cytology TechniqueDeep Vaginal CytologyTwist back and forth, never around and around

40. Bitch will often flag while taking cytology if in estrus

41. Gross Characteristics of the Cytology Swab

42. Preparing the slideAlways roll, Never Smear

43. Stain using fresh stain and sufficient time in each, fix, acidophilic, and basophilic

44. Cytology Results

45. Canine • Artificial Insemination Applications • Inability of breeding animals to travel • Fresh “extended” • Conservation of valuable germ plasm • Frozen semen • Older males do not freeze as well as • Requires surgical insemination or • Transcervical insemination by endoscopy or cannula • Timed by endocrine assay

46. Canine Artificial Insemination • Second successful insemination in any species performed by Spallanzani (1784) • Bitch whelped three pups 62 d later • Technically very simple

47. Canine AI • Equipment • Insemination pipettes • Syringe • Gloves • Artificial vagina or styrofoam cup • Microscope

48. Coital Lock or Tie

49. Assessing Collection: Other considerations • Counts • Depending on size of dog one ejaculate should have 0.3-1.5 billion sperm • Cytology: i.e. are their inflammatory cells etc? • Composition of the ejaculate • Sperm rich fraction • Small in volume and “first” fraction • Pre-sperm fraction is practically speaking, a myth • Prostatic fraction • Last fraction, large volume • Volume may depend on how long one collects for

50. Insemination of the Bitch • Not a sterile procedure • Moderate aseptic technique • Under influence of estrogen, the tract is relatively resistant to infection • Progesterone decreases this resistance • Pyometra