1. Jesus Manuel Palomino DVM, MSc University of Saskatchewan
41st Annual Conference of the IETS, Versailles, France January 10-13, 2015
Superovulation: A tool to produce disease free embryos in wood bison (Bison bison athabascae)
Jesus Manuel Palomino DVM, MSc
PhD Candidate
University of Saskatchewan
Recipient of:
Peter W. Farin Trainee Travel Scholarship Award

2. The Problem Reproductive Germ plasm Disease-free
Hartop et al., 2009
Genetic diversity
Brucellosis and tuberculosis
Disease-free areas
Population decline
Loss of genetic diversity
Reproductive Germ plasm Disease-free
Technologies bio-bank offspring

3. Reproductive technologies to produce disease-free wood bison embryos
In vitro embryo production
Superstimulation and oocyte collection
In-vivo-derived embryos
Superovulation and embryo collection

4. Progression of thesis research
Study 1. Inducing ovulation in Wood Bison
Study 2. Effect of LH vs hCG and progesterone on superovulation
Study 3. Does eCG improve the superovulatory response and embryo quality in wood bison?
Study 4. …
Study 5. …  

5. What did we learn from Studies 1 to 3?
Ovarian superstimulation:
- 8 vs 2 vs 1 administrations of FSH
Induction of ovulation:
- hCG more effective than LH
Adjunct treatments
eCG: Did not improve superovulation or embryo quality
Exogenous progesterone not needed: anovulatory season
Problem
Number of embryos collected: 4 embryos/cow
Number of Grade 1 & 2 (Freezable) embryos: 2 embryos/cow
Can we improve these numbers?

6. Study 4
Effect of lengthening of FSH treatment on superovulation and embryo quality in wood bison
Rationale:
In cattle, lengthening the period of FSH treatment induced a greater number of ovulations and CL compared with conventional treatment (Garcia Guerra et al., 2012)
Lengthening FSH treatment will rescue more subordinate follicles in the wave and they will grow to a sufficient stage of maturity to respond to LH and ovulate

7. Hypothesis
Lengthening FSH treatment will improve the ovulation rate and embryo quality in superovulated wood bison.
Objective:
Determine the effect of lengthening of FSH treatment on superovulatory response and embryo quality in wood bison during the ovulatory and anovulatory season

8. Experiment 1: Anovulatory season
Experimental design:

9. Experiment 2: Ovulatory season
Experimental design:

10. Animals and facilities
Alternative Livestock Center, University of Saskatchewan
50 adult female and 4 male wood bison
Hydraulic chute to handle bison

11. Synchronization: Follicular ablation
Day -1
Day 0

12. FSH 400 mg (Folltropin, Bioniche)
Superovulation
FSH 400 mg (Folltropin, Bioniche)
Hyaluronic acid (Map 5, Bioniche)
hCG 3000 IU (Chorulon, Merck)

13. Artificial Insemination: Chilled semen
Semen collection: electroejaculation, 3 bulls, pooled
Extender: Tryladil (MOFA) + 20% egg yolk
Chilled semen 5º C (1-3 days)

14. Embryo collection: Non-surgical

15. Embryo classification (IETS, 2010)
cvzd

16. Table 1: Anovulatory season
FSH Short
FSH Long
Number of bison 16
Number of follicles ≥9 mm on Day 5
10.8 ± 2.0
10.8 ± 1.1
Ovulation rate
(# ovulations / # follicles ≥9 mm)
52%
(67/130)
54%
(70/130)
Number of CL
4.5 ± 0.9
5.7 ± 1.0
Total ova-embryos
2.6 ± 0.5
3.2 ± 0.7
Grade 1 & 2
0.5 ± 0.3
0.3 ± 0.2
Grade 3
0.1 ± 0.1
Grade 4
1.2 ± 0.4
1.8 ± 0.6
Recovery rate
(# ova-embryos/# CL)
48 %
(26/54)
51%
(35/68)
Results are presented as Mean±SEM, otherwise stated.

17. Table 2: Ovulatory season
FSH Short
FSH Long
P - value
Number of bison 14
Number of follicles ≥9 mm on Day 5
8.1 ± 1.1
9.8 ± 1.4
0.40
Ovulation rate
(# ovulations / # follicles ≥9 mm)
60%
(67/130)
74%
(73/98)
0.07
Number of CL
4.6 ± 0.6
6.5 ± 0.9
0.13
Total ova-embryos
2.2 ± 0.5
4.4 ± 0.7
0.04
Grade 1 & 2
1.1 ± 0.4
2.3 ± 0.6
0.09
Grade 3
0.2 ± 0.1
0.8 ± 0.4
0.03
Grade 4
0.8 ± 0.2
1.7 ± 0.6
0.05
Recovery rate
(# ova-embryos/# CL)
48 %
(22/46)
68%
(44/65)
0.06
Results are presented as Mean±SEM, otherwise stated.
Garcia Guerra et al., 2012
Dias et al., 2013

18. Conclusions
Extended superstimulatory protocol improved embryo production during the ovulatory season
Embryo quality was not affected by lengthening the superstimulatory protocol
Repeat experiment with greater number of bison

19. Production of disease-free embryos from superovulated wood bison
Study 5:
Production of disease-free embryos from superovulated wood bison
Objective:
Determine the effectiveness of washing procedures for removing Brucella abortus from bison embryos exposed in vitro
Abstract 140

20. Level 3 laboratory – Intervac/VIDO

21. Acknowledgments Department of Veterinary Biomedical Sciences Miriam
Dr. Adams
Dr. Mapletoft
Dr. Woodbury
Dr. McCorkell
Dr. Muir/ Dr. Machin
Dr. Anzar
Dr. Mastromonaco
Miriam
Rodrigo, Hayder, Alisha
Nick, Brad
Kosala, Lyle
Department of Veterinary Biomedical Sciences