Implementation of a Successful Embryo Transfer program Brad Stroud, DVM ARSBC 2014 Stillwater, OK Donor Recipient Flush Unfertilized Ova Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos Donor Cow Genetically Superior Female Donates her embryos Donor Recipient Flush Unfertilized Ova Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos Recipient (surrogate) Receives an embryo Donor Recipient Superovulation Unfertilized Ova Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos ARSBC 2014 1
Superovulation - FSH Single ovulating cow Moderate superovulation Heavy Superovulation Donor Recipient Flush Unfertilized Ova Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos Flush: recovering embryos from a superovulated donor female by running fluid in and out of her uterus multiple times. A flush can also be called a collection. Donor Recipient Flush Unfertilized Ova (UFO) Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos Unfertilized Ova Donor Recipient Flush Unfertilized Ova (UFO) Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos ARSBC 2014 2
Degenerate (dead) embryos Donor Recipient Flush Unfertilized Ova (UFO) Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos Viable embryos Donor Recipient Flush Unfertilized Ova (UFO) Degenerate embryos Viable embryos Grade 1 embryos Grade 2 embryos Grade 3 embryos Grade 1 embryo Grade 2 embryo Grade 3 embryo How do we measure success in ET programs? Embryo mass > 85% of total cell mass Embryo mass 50 to 85% of total cell mass Embryo mass < 50% of total cell mass ARSBC 2014 3
Indicators of Success 1. Embryo production 2. ET conception rates 3. Consistency Viable embryos/flush National Average What should you expect? Dairy Donors 6.2 Beef Donors 6.9 2012 AETA Stats Committee Michael Wehrman Percent of total ova recovered that are viable embryos Dairy Donors 57.9% National Average Beef Donors 54.6% 2012 AETA Stats Committee Michael Wehrman 2012 AETA Stats Committee Michael Wehrman Percent viable: 10/16 = 62.5% ARSBC 2014 4
Brad Stroud presentation Oct. 9, 2014 Percent viable: 1/13 = 8% 7 UFOs Percent viable: 0/7 = 0% 2012 AETA Stats Committee Michael Wehrman 2012 AETA Stats Committee Michael Wehrman What can you do to influence the number of viable embryos per flush? Donor Manager 1. 2. 3. 4. Knows his cows and they know him Manages stress on cows Understands forage management Understands and implements FSH protocols and prescribed doses 5. Good heat detector 6. Good AI technician timing and placement 7. Above all manages infertility & minimizes unproductive flushes ARSBC 2014 Proven donor manager Proven ET practitioner Donor selection Donor management Proven donor manager Proven ET practitioner Donor selection Donor management 5
ET Practitioner Proven donor manager Proven ET practitioner Donor selection Donor management 1. Donor Selection 1. Young to middle aged cows for best results 2. Flush wet cows for best results Effect of age on a single superovulation of 633 reproductively-healthy Holstein cattle Age group No. animals Mean no. ova Mean no. good embryos Heifers 28 6.1 3.8 2 6 years 308 10.4 6.7 7 10 years 224 10.6 6.9 11 14 years 64 9.7 5.3 3 year old donor lactating 12 year old donor dry >15 years 9 5.6 2.6 (Hasler et al., 1983) Donor Selection: Age at SVES Client Collections Donors 10 Yrs High 172 2 (<1) Low 63 8 (13) Viable Emb 1681 (9.8) 283 (4.5) 1. Donor Selection 1. Virgin beef heifers should be 14 months old 18 month old heifer 10 month old heifer ARSBC 2014 6
Brad Stroud presentation Oct. 9, 2014 Heifer Age at Flush Lactating/Dry Donor Data Beef Heifers SVES SVES Data (Multiparous Beef Females) Viable Embryos per Collection Viable Emb/Collection < 14 Mo (n=97) = 3.4 14 Mo (n=266) = 5.2 Dry 6.3 Wet 8.1 Donor Selection Antral follicle count pre superovulation 1. Donor Selection 1. Good flush history 24 good embs 8 good embs 7 good embs Low antral follicle count 1 viable emb 5 viable embs 0 embs High antral follicle count Antral Follicle Count correlates with AMH levels 1. Donor Selection 1. Moderate flesh not thin or fat ARSBC 2014 Proven donor manager Proven ET practitioner Donor selection Donor management 7
Brad Stroud presentation Oct. 9, 2014 Good heat detection systems in place 1. Donor management control # of negative flushes Avoid over-flushing. Flush donors 2 or 3 times between calves. Donor management control # of negative flushes Don t let dry donors get too fat Does selection and management work? SVES Donor Collection Data 2002-2005 (Beef Cattle) Evaluate frozen semen in advance of inseminating donor Client Collections Total Ova Viable Emb Per flush % of Ova Viable High 172 2719 (15.8) 1681 (9.8) 63.4 Low 63 593 (9.4) 283 (4.5) 48.2..\..\..\..\Video Clips\Semen\Tutorial Video Files\Semen DIC 10x Linear Spiraling and medium speed DVD.mpg..\..\..\..\Video Clips\Semen\Tutorial Video Files\Semen DIC 10x 10 perc Mot mrd and tite circles DVD.mpg...\..\..\..\Video Clips\Semen\Phase C\Dead Sperm\Semen Dead.mpg ARSBC 2014 Farms with minimum of 50 collections 8
Brad Stroud presentation Oct. 9, 2014 Collections with zero viable embryos High Client Low Client Total Collections 172 63 Collections with 0 viable 7 (4) 12 (19) Recipient selection = high or low preg rates Most common mistake in recipient management is failure to cull old dry fat cows from herd 1. Embryo production 2. ET conception rates and/or # offspring produced 3. Consistency Most common mistake in recipient selection is old dry fat cows from within herd Manage recipients at time of transfer for highest preg rates Cull open recipients after 2 good chances, i.e. 2 grade one embryos with good synchrony ARSBC 2014 9
Resources (man and machines) are important for high volume throughput. Restraint of recipient during transfer is crucial To achieve consistent results with superovulation and ET requires knowledge, experience, resources, and skill sets on many fronts. Any weaknesses in a program can lead to less than desired results ARSBC 2014 10