1. The Estrous Cycle Physiology and Endocrinology

2. Terminology Estrus is a noun. –The cow is displaying estrus. Estrous is an adjective. –The length of the estrous cycle is 21 days. Oestrus and Oestrous –British and European spellings Estrus and Heat are synonymous

3. Terminology (cont.) Anestrus –When the female is not having repeated estrous cycles. Polyestrus –repeated estrous cycles Seasonally Polyestrus –repeated estrous cycles only certain times of the year Monoestrus –only one cycle per year

4. Average Reproductive Cycles Species Length of Length ofOvulation Length of Estrous Cycle EstrusPregnancy cow 21 days 18 hr11 hr after282 days polyestrusend estrus ewe 17 days 29 hrnear end148 days seasonal (fall)estrus sow 21 days48-72 hr35-45 hr115 days polyestrusafter start estrus mare 21 days4-8 days3-6 day of335 days seasonal (spring)estrus polyestrus(1-2 days before end of estrus)

5. Variation in Cycle Types ExampleType of Follicular Ovulation & CL CycleDevelopmentCL Formation Function Cow, ewe, LongSpontaneousSpontaneousSpontaneous sow, mare rats, mice, ShortSpontaneousSpontaneous Induced hamsters (4 days) (prolactin) rabbit, cat,InducedSpontaneous Induced Induced mink, ferret, (LH surge) otter, alpaca

6. Estrous Cycle  Length of the estrous cycle  Average 21 days (range 18 to 24 days)  Estrus (standing heat)  12 to 18 hours (range 8 to 30 hours)  Ovulation  Approximately 30 hours after the beginning of standing heat (or 12 to 18 hours after the end of standing heat) 2

7. Estrous Cycle  Major structures on the ovary are …  Follicle … a blister-like structure containing the egg (referred to as oocyte); produces hormone “estrogen”  High amount of estrogen causes “standing heat” and “ovulation”  Corpus luteum (referred to as “CL”) … looks like a hard yellow structure and produces hormone “progesterone” that is responsible for maintenance of pregnancy 3

8. Phases in oestrous cycle 1.Proestrus 2.Estrus 3.Metestrus 4.Diestrus

9. Follicular Phase Proestrus follicle enlarges estrogen increases vascularity of the female reproductive tract increases endometrial glands begin to grow estrogen levels peak

10. Estrus allows male to mount estrogen decreases LH surge occurs ovulation 24-48 hr after surge of LH uterine motility high with contractions moving toward oviduct sperm transport is optimal cervical mucus volume increases Follicular Phase

11. Luteal Phase Metestrus estrogen low ovulation in cow corpus hemorrhagicum present uterus  contractions subside  endometrial glands continue to grow and become coiled  in cattle bleeding occurs FSH increases  triggering growth of follicles

12. Luteal Phase Diestrus progesterone high FSH  Increases at some point to cause growth of ovulatory follicle Uterus  secrets fluid but volume gradually decreases  contraction stop  CL regresses at the end of this period if female is not pregnant due to PGF release

13. Characteristics of Estrous Cycles Cow Ewe Sow Mare Estrous cycle (days) 21 17 21 21 Proestrus (days) 3-4 2-3 3-4 2-3 Estrus12-18 hr24-36 hr48-72 hr4-8 days Metestrus (days) 3-4 2-3 2-3 2-3 Diestrus (days)10-1410-12 11-13 10-12

14. Estrus phase is the shortest phase in the estrous cycle, but the most important one….!! Why….?

15. Follicular Development During Estous Cycle

17. Ovarian Structure

19. Follicle Development Follicular Wave Recruitment Growth Selection Dominance Regression Next Recruitment  Follicle development occurs as a wave-like pattern consisting of “Recruitment”, “Selection”, “Growth”, “Dominance”, and “Regression” phases  Usually 2 to 4 follicular waves occur during the estrous cycle in cattle 4

20. Follicle Development Follicle Stimulating Hormone (FSH) Recruitment Next Recruitment  FSH precedes recruitment of follicles (causes follicles to start growing)  FSH is the same hormone used for superovulation and embryo transfer in cattle 5 Regression FSH

21. Follicle Development Luteinizing Hormone (LH) Growth Selection Dominance  LH promotes further follicle growth and maturation of egg 6 Regression LH pulses

22. Follicle Development and Ovulation Luteinizing Hormone (LH) and Estrogen Growth Selection Dominance Estrus LH Surge Ovulation Estrogen  LH stimulates follicle growth and a growing follicle produces high levels of estrogen  High levels of estrogen, in turn, cause estrus and surge release of LH that triggers ovulation 7

23. 024681012141618200 Estrus Day of the Estrous Cycle  Length of the estrous cycle in cattle with 3 follicular waves is typically 20 to 24 days Follicle Development During the Estrous Cycle Example for 3 Follicular Waves 8 Ovulation

24. Day of the Estrous Cycle 024681012141618200 Estrus  Length of the estrous cycle in cattle with 2 follicular waves is typically 18 to 20 days, slightly shorter than the estrous cycle with 3 follicular waves 9 Ovulation Follicle Development During the Estrous Cycle Example for 2 Follicular Waves

25. 024681012141618200 Estrus Day of the Estrous Cycle 10 Ovulation Endocrinology During the Estrous Cycle Follicle Stimulating Hormone (FSH)  Example shown for cattle having 3 follicular waves during a 21-day estrous cycle FSH

26. 024681012141618200 Estrus Day of the Estrous Cycle  Example shown for cattle having 3 follicular waves during a 21-day estrous cycle 11 Endocrinology During the Estrous Cycle Luteinizing Hormone (LH) LH Surge Ovulation LH Surge LH pulses

27. 024681012141618200 Estrus Day of the Estrous Cycle 12  Example shown for cattle having 3 follicular waves during a 21-day estrous cycle Endocrinology During the Estrous Cycle Estrogen Ovulation Estrogen

28. 024681012141618200 Estrus Day of the Estrous Cycle Growth Regression 13 Corpus Luteum (CL) Growth and Regression  Corpus luteum develops from the ovulated follicle and takes approximately 10 days to reach mature size Ovulation

29. 024681012141618200 Estrus Day of the Estrous Cycle  Corpus luteum produces progesterone  Progesterone is responsible for maintenance of pregnancy after conception occurs 14 Corpus Luteum (CL) Progesterone GrowthRegression Progesterone

30. 024681012141618200 Estrus Corpus Luteum (CL) Regression Prostaglandin F 2  (PG) Day of the Estrous Cycle  Late in the estrous cycle, uterus produces PG which causes regression of corpus luteum  PG is the same or similar hormone in “Lutalyse ® ”, “Estrumate ® ”, “ProstaMate ® ”, and “In Synch ® ” GrowthRegression 15 PG

31. Estrus Corpus Luteum (CL) Maintenance When cow becomes pregnant … Day of the Estrous Cycle  Presence of embryo blocks uterus to produce PG late in the estrous cycle which causes maintenance of corpus luteum and production of progesterone for pregnancy PG Maintenance 024681012141618200 Embryo Growth 16

32. Progesterone regulates LH Pulses Luteinizing Hormone (LH) 024681012141618200 Estrus Day of the Estrous Cycle LH Surge  Progesterone regulates secretion pattern of LH pulses and hence, follicular development 17 Progesterone LH pulses

33. Endocrinology of the Estrous Cycle 024681012141618200 Estrus Day of the Estrous Cycle Estrogen Progesterone PG  Relationships among estrogen, progesterone, and PG during the 21-day estrous cycle 18

34. Physiology and Endocrinology of the Estrous Cycle 024681012141618200 Estrus Day of the Estrous Cycle 19  Relationships among structural and hormonal changes during the 21-day estrous cycle (example shown for cattle having 3 follicular waves)

35. Time to breed LABORATORIUM REPRODUKSI TERNAK FAPET UB 35

36. Causes of Anestrus Pregnancy Presence of Offspring Season Stress Pathology Nutrition

37. Gestational Anestrus Progesterone during pregnancy –negative feedback After parturition anestrus continues –progesterone exposure during pregnancy –hypothalamus Lacks estradiol positive feedback allows time for uterine involution

38. Seasonal Anestrus just like entering puberty silent ovulation

39. Silent Ovulation Anestrus Estrus

40. Lactational Anestrus

41. Suckling Other offspring factors –visual encounter –olfactory encounter –auditory encounter Prolactin –inhibits GnRH release –major infertility problem in women who are not lactating

42. Nutritional Anestrus

43. Terima kasih…

45. Estrus Synchronization 20

46. Effective Estrus Synchronization Programs  Shorten the AI breeding season  Cows or heifers are in estrus during a predictable interval that facilitates AI  Reduce time and labor required to detect estrus  Result in more cows and heifers becoming pregnant early during the breeding season  Progestin-based programs can induce estrous cyclicity in anestrous cows and prepubertal heifers (For example: MGA, or CIDR)* 21 * Progestins are progesterone-like compounds that act like progesterone

47. Effective Estrus Synchronization Programs  Result in older and heavier calves at weaning  Will have beneficial effects on the next breeding season  More cows and heifers calve early  More days postpartum at the next breeding season  Replacement heifers will be older 22

48. Effective Estrus Synchronization Programs  Consider what happens during a restricted breeding season, based on the average 21-day estrous cycle:  If cows or heifers are cycling when an estrus synchronization treatment is implemented and they exhibit estrus during the synchronized period, they would have 3 opportunities to conceive during a 45-day period or 4 opportunities during a 65-day period  If cows or heifers are cycling but no estrus synchronization treatment is implemented, then they have only 2 (45-day) or 3 (65-day) opportunities to conceive  If cows or heifers are not cycling at the beginning of the breeding season, they have even less opportunity to conceive 23

49. Points to Consider When Using Estrus Synchronization  Estrus synchronization is never a substitute for:  Nutrition  Herd health  Proper management  Estrus synchronization should not be used as a crutch for poor management  When administered appropriately, estrus synchronization is an effective reproductive management tool that can be used to facilitate AI 24

50.  Determine which females would make the best candidates before beginning estrus synchronization  Successful application of estrus synchronization is easier to accomplish with heifers (no calves to work with) Points to Consider When Using Estrus Synchronization 25

51. Selecting Heifers for Estrus Synchronization  Heifers should be of adequate age (cycling or close to reaching puberty)  Heifers should be of adequate weight  Developed to 65 % of projected mature weight  Do you know the mature weight of your cows?  Utilize reproductive tract score (RTS) to assess reproductive maturity 37

52. Timing of Puberty in Heifers  Target weight: Heifers reach puberty at approximately 65% of their mature body weight  Determine your desired weight at breeding, calculate the gain needed to meet that weight, and feed to meet it 38

53. Management Considerations for Heifers  DO NOT use growth promoting implants in replacement heifers  Implants may disrupt or impair normal development of reproductive organs  Developing uterus is especially sensitive to growth promoting implants 39

54. Management Considerations for Heifers  Reproductive Tract Scores (RTS) are determined by rectal palpation of the ovaries and uterus  The RTS should be performed approximately 6 to 8 weeks prior to breeding season and/or approximately 2 weeks prior to beginning an estrus synchronization treatment 40

55. Reproductive Tract Scores (RTS) RTS 1 2 3 4 5 Uterine horns Immature < 20 mm diameter No tone 20-25 mm diameter No tone 20-25 mm diameter Slight tone 30 mm diameter Good tone > 30 mm diameter Length 15 18 22 30 > 32 Height 10 12 15 16 20 Width 8 10 10 12 15 Ovarian Structures No palpable follicles 8 mm follicles 8-10 mm follicles > 10 mm follicles CL possible CL present Ovarian Measurement (mm) Adapted from Anderson et al., 1991 Description Infantile Prepubertal (more than 30 days to puberty) Peripubertal (within 30 days to puberty) Cycling (Follicular Phase) Cycling (Luteal Phase) 41

56. RTS Measurements RTS 1 2 3 4 5 No. of Heifers 61 278 1103 494 728 Weight (lb) 594 a 620 b 697 c 733 d 755 d Pelvic Width (cm) 10.9 a 11.2 a 11.4 b 11.7 c Pelvic Height (cm) 13.9 a 14.1 a 14.5 b 14.7 c Pelvic Area (cm 2 ) 152 a 158 a 166 b 172 c Estrous Response 54 % a 66 % b 76 % c 83 % d 86 % d Adapted from Patterson et al., 1999a 42  Note the relationship between increasing RTS, weight, and skeletal development determined by pelvic area  Estrous response increased among heifers that were more reproductively mature at the beginning of estrus synchronization and/or breeding season a, b, c, d = P < 0.05

57. Relationship Between RTS and Pregnancy RTS 1 2 3 4 5 No. of Heifers 53 762 3458 3398 2613 Pregnant 31 610 3014 3006 2331 Open 22 152 444 392 282 Pregnancy Rate 58 % 80 % 87 % 89 % Randle et al., 2001 43  Higher pregnancy rates at the end of the breeding season were achieved among heifers that were more reproductively mature at the beginning of estrus synchronization and/or breeding season

58. Prepubertal Anestrus  Timing of puberty is controlled by …  Age  Weight  Breed  Season  Develop heifers to approximately 65% of mature weight by the breeding season  This will help them to attain puberty at the right time 53 Many factors involved

59. 55 Nutritional Anestrus  Nutritionally stressed cattle will experience delays in return to estrus after calving  Can be “managed” through  Improvement in BODY CONDITION  Proper feeding management

60. 56 Postpartum Anestrus Induction of Estrous Cyclicity  Bull exposure  Prepubertal heifers Exposure of prepubertal heifers to sterile bulls (for example, vasectomized, epididyectomized … etc) for 60 to 80 days before the breeding season may hasten the onset of puberty (ranges from no effect to 70 days earlier)  This procedure also shortens the postpartum period in anestrous cows however my concentration is on heifers Izard and Vandenbergh, 1982; Berardinelli et al., 1978; MacMillan et al., 1979; Roberson et al., 1991 Foote, 1974; MacMillan et al., 1979; Zalesky et al., 1984; Gifford et al., 1989

61. Products Currently Utilized in Protocols to Synchronize Estrus *** Price may vary depending on suppliers ***  Prostaglandin F 2  (PG) Lutalyse ®, Estrumate ®, ProstaMate ®, In Synch ®  Approximate cost is $3.00/dose (range $2.50-$4.00)  Gonadotropin-Releasing Hormone (GnRH) Cystorelin ®, Factrel ®, Fertagyl ®  Approximate cost is $5.00/dose (range $4.00-$6.00)  Melengestrol Acetate (MGA ® Premix)  Approximate cost ranges $0.14 - $0.40/day/head including carrier  14 days ($2.00-$5.60/head) or 7 days ($1.00-$2.80/head)  EAZI-BREED™ CIDR ® Cattle Insert (CIDR)  Approximate cost is $8.00 (range $8.00-$10.00)  Syncro-Mate-B ® (off the market) 28

62. Prostaglandin F 2  (PG) 60

63.  PG causes CL regression  No effect on anestrous cattle  No induction of estrous cyclicity … No Jump-start  Only effective during Days 6 (6 days after estrus) to 16 of the estrous cycle Prostaglandin F 2  (PG) 61 Lauderdale, 1972

64. 024681012141618200 Day of the Estrous Cycle PG regresses CL NO YES Prostaglandin F 2  (PG) Estrus 62

65. Modified from Stevenson et al., 2000 Hours following second PG injection % of cows in estrus 0 10 20 30 40 50 0< 24364860728496108120> 120 PG 2 x PG 77 Estrous Response Double Injection PG Program Example shown is 14-day interval for PG injections

66.  Advantages:  Cost effective ($3.00 or $ 6.00)  Easy to use  Good fertility Single or Double Injection PG Programs $3.00 or $6.00 ($3.00/one dose) 74

67.  Disadvantages:  No effect between days 0 to 4 or 16 to 21 of the estrous cycle  Improved effectiveness after day 10 of the estrous cycle  Require heifers and cows to have CL (cycling)  Require extensive estrus detection (minimum 7 days)  No induction of estrous cyclicity (No Jump-start) Single or Double Injection PG Programs $3.00 or $6.00 ($3.00/one dose) 75

68.  Estrous response:  Varies depending on the degree of estrous cyclicity in the herd and the stage of the estrous cycle of an individual animal  Single injection of PG … up to 40 to 60 %  Double injection of PG … up to 50 to 80 %  Timing of estrus:  Before PG injection to 7 to 10 days after PG injection  Peak response ranges from 48 to 96 hours after PG injection Possible Outcomes Single or Double Injection PG Programs 76

69. Melengestrol Acetate (MGA ® ) 98 MGA is a registered trademark of Pfizer Animal Health

70.  Orally active progestin, administered as a feed additive  Blocks estrus and ovulation  Acts similar to progesterone from CL  MGA can induce estrous cyclicity in peripubertal (within 30 days to attain puberty) heifers and postpartum anestrous cows  Currently, MGA and CIDR are the only progestins commercially available in the U.S. Melengestrol Acetate (MGA) 99 Patterson et al., 1989

71.  Mix with grain source (example: cracked or ground corn) For example: MGA with ground corn ($0.15/day/head)  Top dressing does not work well  If cattle are on lush pasture, remove salt from pasture and include salt in MGA carrier 0.5 oz. Salt per cow and MGA in carrier ($0.30/day/head)  Pellet form (very palatable) For example: MFA Cattle Charge ® with MGA ($0.40/day/head) Intake Considerations What type of carrier to use? 100

72.  Need to provide adequate bunk space (18 inches per heifer, and 24 inches per cow)  Make sure cattle come to bunk before start of feeding MGA  If cattle are not used to bunk feeding, 1-2 weeks of warm-up bunk feeding is recommended  MGA should be fed one time per day at approximately the same time every day  Separate heifers from cows for MGA feeding  Late-term pregnant cows should not be fed MGA Intake Considerations 101

73. MGA can be combined with PG to improve results

74. Distribution of Estrus MGA-PG (for AI) MGA only (for Natural service) % of herd in estrus  Adding the PG injection to the MGA program results in a much tighter synchronization of estrus 3-5 days 7-10 days 106 Adapted from Patterson et al., 2000

75. Management Considerations Why Not Introduce Bulls Right After MGA Withdrawal? 0 5 10 15 20 25 30 Days after MGA feeding % of herd in estrus  If bulls were introduced to the herd right after MGA withdrawal, they may be overworked during the less fertile estrous period after MGA withdrawal, but before cows express fertile estrus Less fertile estrus after MGA 7-10 days 107 2 nd estrus for anestrous cows after short estrous cycle (more fertile) 2 nd estrus for most of cows (more fertile) Suggested timing of introducing bulls (see page 104 and 105)

76.  Advantages:  Easy to use  Shortens the next calving season (ideal way to start AI program in the following year)  MGA can induce estrous cyclicity in postpartum anestrous cows (Jump-start)  Does not require cattle to be handled Natural Service with MGA Program $2.00–$5.60 (MGA cost) 109

77.  Disadvantages:  Must have adequate bull-power  At least 1:15 to 20 bull:female ratio for 2-year or older bulls  Yearling bulls … consider decreasing the bull:female ratio  Resulting calves may be of inferior quality and provide less genetic improvement for the herd compared to AI calves (depends on the genetic quality of the clean-up bulls) Natural Service with MGA Program $2.00–$5.60 (MGA cost) 110

78. Summation Lots of programs –Many combinations of programs Many prices The numbers definitely show results –Earlier calving season –More time to return to Estrus –Increased opportunity to become pregnant However due to expense and labor intensity this is not for everyone

79. Bibliography “Guide to Estrus Synchronization of Beef Cattle” CD Freddie N. Kojima, Ph.D.and David J. Patterson, Ph.D. Department of Animal Sciences University of Missouri Columbia, MO 65211